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NB100-2220

LC3B Antibody - BSA Free

(51 Reviews)

(1861 Publications)

0.1 ml

NB100-2220

Lysates of HeLa parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50 uM CQ for 18 hours. PVDF (Polyvinylidene difluoride) membrane was probed with 0.5 ug/mL of Rabbit Anti-LC3B Polyclonal Antibody (Catalog # NB100-2220) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog# HAF008). A specific band was detected for LC3B at a molecular weight of approximately 15 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH is shown as a loading control. This experiment was conducted under reducing conditions.

<b>Genetic Strategies Validation. </b>LC3B was detected in immersion fixed CQ treated HeLa cells (left) but was not detected in LC3B knockout HeLa cells (right) using rabbit anti-human LC3B polyclonal antibody (Catalog #NB100-2220) at 0.3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights(TM) 557-conjugated anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm.

<b>Biological Strategies Validation. </b>LC3B-Antibody-Western-Blot-NB100-2220-img0073.jpg

LC3B-Antibody-Immunohistochemistry-NB100-2220-img0074.jpg

LC3 accumulation in muscle fibers from patients with PAD (row B - E). Normal fibers from a Non-PAD sample (row A). Scale bar = 100 uM. Image from verified customer review.

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Detection of LC3B in mouse ES cell lysates using Rabbit anti- [Catalog # NB100-2220]. LC3 detected at a molecular weight of approximately 17 kDa. Atg5-/- ES cells from Dr. Noboru Mizushima [Mizushima, N. et al. J. Cell Biol. 152 (2001)] Photo courtesy of Dr. Beth Levine, UT SW Medical Center.

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<b>Biological Strategies Validation. </b>Lysates of mouse NIH3T3 and rat PC-12 cell lines untreated (-) or treated (+) with CQ. PVDF (Polyvinylidene difluoride) membrane was probed with 0.5 ug/mL rabbit anti-LC3B polyclonal Antibody (Catalog # NB100-2220, Novus Biologicals), followed by 1:2000 dilution of goat anti-rabbit IgG secondary antibody. LC3 detected at a molecular weight of approximately 15 kDa in treated NIH3T3 and PC-12 cells.

<b>Biological Strategies Validation. </b>Detection of LC3I and LC3II in mouse cochlea cell line SV-K1 using Rabbit anti- [Catalog # NB100-2220]. Cells were treated with CQ (1 uM), and As2O3 (1 uM) for 24 hrs. Image from verified customer review.

<b>Biological Strategies Validation. </b>Autophagy Investigation. (A) Representative WBs (Western Blots) of LC3B-I, LC3B-II and GAPDH loading control in DANs derived from 4 iPD patients and 4 healthy controls. (B) WBs from three independent experiments were quantified by densitometry and LC3B-II normalized to a loading control. Grouping of data by healthy vs iPD as well as iPD stratified for T/T and C/C genotype. (C) Ratio of LC3B-II (normalized to loading control) following bafilomycin treatment/untreated. Citation: Marrone L, Bus C, Schondorf D, Fitzgerald JC, Kubler M, Schmid B, et al. (2018) Generation of iPSCs carrying a common LRRK2 risk allele for in vitro modeling of idiopathic Parkinson's disease. PLoS ONE 13(3): e0192497. //doi.org/10.1371/journal.pone.0192497

<b>Biological Strategies Validation. </b>LC3B-Antibody-Western-Blot-NB100-2220-img0067.jpg

<b>Biological Strategies Validation. </b>LC3B-Antibody-Western-Blot-NB100-2220-img0071.jpg

LC3B-Antibody-Immunocytochemistry-Immunofluorescence-NB100-2220-img0064.jpg

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Staining of rat brain tissue section. Analysis using Rabbit anti-LC3B Antibody [Catalog # NB100-2220]. Image from verified customer review.

Human ovarian Cancer tissue stained using heat mediated antigen retrieval in pH 6.0 citrate buffer at 1:200 dilution. Image provided by verified customer review.

FFPE (Formalin-Fixed Paraffin-Embedded) tissue section of mouse brain using 1:200 dilution of Rabbit anti-LC3B antibody [Catalog # NB100-2220]. The specific signal of LC3 was detected using HRP-conjugated secondary antibody with DAB (3, 3 -diaminobenzidine) reagent, and nuclei of cells were counterstained using hematoxylin. This LC3B antibody generated a low to moderate levels of cytoplasmic staining in the glial cells. The neurons depicted a moderate to strong staining for LC3 in their cytoplasm.

Analysis using the Biotin conjugate of Rabbit anti-LC3B Antibody [Catalog # NB100-2220]. Staining of brain, cerebral cortex, neurons with cell processes.

Image shows a specific band for LC3B at a molecular weight of approximately 15 kDa in 0.5 mg/mL of Neuro2A lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

TP53 knockout triggers a pro-mitophagic response.PINK1 (a,b, N = 8), LC3-2/LC3-1 (a,c, N = 8), p62 (a,d, N = 12), TIM23 (a,e, N = 10), TOM20 (a,f, N = 9), HSP60 (a,g, N = 16), optineurin (a,h, N = 9) & NDP52 (a,i, N = 8) protein levels were analyzed in control (p53+/+) or TP53-deficient (p53−/−) HAP1 cells as described in the Methods. Bars represent the means ± SEM of 3-4 independent experiments performed in triplicate & are expressed as percent of control (p53+/+) cells. Actin expression is provided as a representative gel loading control in a. Statistical analyses were performed with GraphPad Prism software by using unpaired Student’s t-test. Significant differences are: ** p 50%; & staining (E) absent, (F) weak, (G) moderate, (H) strong. (upper panel, x200; lower panel, x400). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29190884), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - P53 mediates topotecan-induced autophagy through the activation of sestrin 2 & AMPK in colon cancer cells with wild-type p53.A. HCT116 & LS-174T cells were treated with various concentrations of TPT for 24 h. The levels of P53, sestrin2 & p-AMPK were analysed by immunoblotting. B. HCT116 cells were transfected with p53 or sestrin 2 siRNAs for 24 h, treated with or without 1 µg/mL TPT for an additional 24 h, & the indicated proteins were then analysed by immunoblotting. C. HCT116 cells were transfected with LKB1 or CaMKK beta siRNAs, treated with or without 1 µg/mL TPT for an additional 24 h, & the indicated proteins were detected by immunoblotting. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23024792), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Specific inhibitor SP600125 or siRNA directed JNK blocked ceramide-induced autophagy & up-regulation of LC3 expression. (A) SUNE1 cells were treated with 20 μM ceramide for 24 h in the absence or presence of SP600125 or JNK1/2 siRNA. Lysates were analyzed by immunoblotting. (B) Autophagosome formation was visualized using YFP-LC3 expressing & observed under a confocal microscope. Representative immunofluorescence pictures are shown at the original magnification × 1000. (C) The expression of LC3 mRNA was examined by RT-PCR analysis. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21943220), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Glutamine deprivation induces IL-8 secretion.(a) RT–PCR analysis of LC3, IL-8, CHOP & GADD34 in U2OS cells subjected to glutamine deprivation (−Q) or exposed to TPG (1 μM). (b) Immunoblot analysis of ER stress- & autophagy-related proteins in U2OS cells subjected to glutamine deprivation or TPG treatment. Samples are run in triplicate. (c) Cytokine array analysis of conditioned media from U2OS & A549 cells grown in the presence (+Q) or absence (−Q) of glutamine. (d) IL-8 enzyme-linked immunosorbent assay of conditioned media from U2OS & A549 cells grown in the presence (+Q) or absence (−Q) of glutamine. Error bars in all figures represent s.d. of three biological replicates. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms5900), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - NDs API improves ATO-based therapy in vivo. HepG2 tumor-bearing nude mice administered intravenously (i.v.) w/ NS (200 μL), NDs (5 mg kg−1), ATO (0.75 mg kg−1), ATO (1.5 mg kg−1), NDs–0.75 mg kg−1 ATO mixture,/NDs–1.5 mg kg−1 ATO mixture daily for 5 consecutive days every week, w/ 13-week cycles c Immunoblots of autophagy-related proteins LC3-II, p62 (left); semi-quantified analysis (n = 3) in tumor tissues from mice after treatment (right). GAPDH used as loading control. Normalized band densities shown below each band. Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/30341298), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - NDs API improves the ATO-based therapy in vivo. HepG2 tumor-bearing nude mice administered intravenously (i.v.) w/ NS (200 μL), NDs (5 mg kg−1), ATO (0.75 mg kg−1), ATO (1.5 mg kg−1), NDs–0.75 mg kg−1 ATO mixture,/NDs–1.5 mg kg−1 ATO mixture daily for 5 consecutive days every week, w/ 13-week cycles. g Immunostaining of LC3 & ATG13 in tumor tissues of NDs–1.5 mg kg−1 ATO-treated mice. Schematic showing (left) & imaging (right) indicate that LC3/Atg13 puncta in periphery & around the blood vessels inside the tumors. Scale bars: 20 μm. Error bars s.d. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30341298), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: LC3B Antibody - BSA Free [NB100-2220] - LC3, CRF, & HIF-1 alpha expression. (a) for each antibody the spatial distribution of immunohistochemical staining was the same in cases with spontaneous & induced labor. (b) The analysis of data from optical density values of Western blotting bands shows no differences of LC3-II & HIF-1 alpha expression between groups. On the contrary CRF is higher in spontaneous labor. LC3-II, HIF-1 alpha , & CRF are normalized onto alpha -tubulin. (c) No correlation between LC3-II & CRF expression was found. SP: spontaneous labor; PG: induction with prostaglandin; PO: induction with prostaglandin & oxytocin; OX: induction with oxytocin. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23956998), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - The time-dependent effects of melatonin on the LC3-II & Beclin 1 expression in U87MG (a, b) & A172 (c, d) cells. The cells were treated with 1 mM melatonin for 4, 12, 24 or 36 h. PBS was used as a negative control, & rapamycin (Rap 200 nM) served as a positive control. The levels of LC3-II & Beclin-1 expression were determined by a Western blot analysis. The protein bands for each regimen were quantified by densitometry, & their differences are presented in the graph. The values represent the mean ± S.E.M. for three separate determinations. Statistical significance * at P 10 puncta per cell from three independent experiments is depicted. Bars represent mean±s.d. from three independent experiments (>50 cells per experiment). The statistical significance (P value) was determined by a two-tailed, paired Student’s t-test. *P<0.05. (c) Immunoblot analysis of U2OS cells subjected to glutamine deprivation with or without 400 nM BafA1. Cells were pretreated with BafA1 for 1 h before & during exposure to glutamine-deficient medium. Autophagic activity was monitored by detection of p62 & LC3-II proteins. (d) U2OS mCherry-GFP-LC3 cells were cultured in the presence (+Q) or absence (−Q) of glutamine for 18 h. Red vesicles denote autolysosomes, whereas yellow vesicles represent autophagosomes. Bars indicate numbers of yellow vesicles (autophagosomes) or red vesicles (autolysosomes) per cell±s.d. (e) Images of U2OS mCherry-GFP-LC3 cells cultured for 18 h in the presence (+Q) or absence (−Q) of glutamine. Scale bar, 10 μm. (f) Phagophore formation in mCherry-ULK1. Scale bar, 10 μm. (g) mCherry-ATG5-expressing U2OS cells after 24 h in the presence (+Q) or absence (−Q) of glutamine. Scale bar, 10 μm. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms5900), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Glucocorticoids promote BCG survival in macrophages by inhibiting autophagy. (a) RAW264.7 cells were pretreated with dexamethasone (DEX) (1 μM) or vehicle control ethanol (mock) for 6 hr & then challenged with M. bovis BCG (MOI 5) or treated with the vehicle control DMSO or rapamycin (4 μM). LC3 level was detected with Western blot analysis. & GAPDH was used as the loading control. Full-length blots are presented in Supplementary Fig. 12. Data are representative of three independent experiments with similar results. (b,c) RAW264.7 cells were treated as described in (a), & viable bacilli were determined by CFU assays at 6 hpi (b). Survival rate was calculated compared with that of mock-treated cells (c). Data are shown as the mean ± SEM of three independent experiments. *p < 0.05. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-01174-9), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - CysC regulates AMPK activity during the induction of autophagy. (a) Immunoblotting analysis of autophagy regulators. N2a cells expressing SOD1 treated with or without CysC (1 μM) for 6 h. The lysates analyzed by immunoblotting using antibodies for phosphorylated AMPK (pAMPK), phosphorylated mTOR (pmTOR), phosphorylated PKCδ (pPKCδ), Myc & beta -actin. (b) Inactivation of AMPK by mutant SOD1 expression. Each relative pAMPK level normalized by beta -actin in (a) is quantified. *P<0.05, **P<0.01 versus mock. (c) Activation of AMPK by CysC treatment. Relative levels of pAMPK for CysC-treated samples normalized by that of PBS-treated control, which is shown as the broken line, in (a) quantified. *P<0.05 versus PBS-treated controls. (d) CysC induced autophagy through the AMPK activation. N2a cells treated with CysC (1 μM), CC (5 μM) or AICA-riboside (AICAR, 5 mM) for 12 h. The lysates analyzed by immunoblotting using antibodies against pAMPK, LC3 & Tubulin (left panel). Quantification of immunoblots was plotted (right panel). (e) The effect of pAMPK activation on neuroprotection by CysC. N2a cells expressing G85R SOD1 mutant treated with CysC (0.2 μM), CC (5 μM) or AICAR (5 mM). Cell viability was measured by the MTS assay. Data expressed as means±S.E.M. from 3 independent experiments. Each experiment was performed in triplicate. *P<0.01 compared to non-treated control, #P<0.01 compared to CysC-treated one. (f) Immunoblotting analysis of pAMPK in SOD1 transgenic mouse spinal cords. The spinal cord lysates from the transgenic mice at indicated ages analyzed for the levels of pAMPK, SOD1 & beta -actin (upper panel). Expression levels of pAMPK normalized by beta -actin (lower panel). All data expressed as means±S.E.M. from 3 independent experiments Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014459), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - Vps34 inhibition blocks autophagy initiation & causes accumulation of ubiquitin-positive & p62-positive structures. a Representative confocal images of cortical neurons treated with vehicle, Bafilomycin A1 (BafA1) at 50 nM, VPS34IN1 at 3 µM or cotreated for 3 h. Arrows highlight LC3 & p62 structures. Right panel, bar graphs denote average object intensity, per cell (mean ± SEM, N = 49-60 cells, from three independent experiments). Scale bar, 10 µm. ***p < 0.001 in one-way ANOVA, Holm–Sidak’s multiple comparisons test. b Representative confocal images of cortical neurons treated as in a & immunostained for LAMP-1, LC3, & p62. Airyscan insets highlight position of LC3 & p62 structures relative to LAMP-1-positive membranes. Scale bar, 10 µm. c Representative confocal images of cultured cortical neurons treated with vehicle or VPS34IN1 at 3 µM for 24 h. Arrows highlight p62 & ubiquitin colocalization. Scale bar, 10 µm Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-017-02533-w), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - ORP1L & VAP-A form cholesterol-dependent ER-AV contact sites(a) MelJuSo cells expressing GFP or GFP-tagged ORP1L mutants were fixed & stained for LC3 & Calnexin. Y477 & D478 in the FFAT motif were mutated to alanines (A) in the ORP1L ydaa mutant. Scale bar, 10 μm. Right: co-immunoprecipitation for ORP1L (mutants) with VAP-A. GFP-ORP1L mutants or GFP were isolated from lysates of HEK293T cells co-overexpressing HA-VAPA using GFP-Trap beads. Western blot filters were probed for isolated GFP-tagged proteins, the associated HA-VAP-A & the input HA-VAP-A, as indicated. (b) Cryo-immuno-EM on HeLa cells expressing HA-LC3 & GFP-ORP1L delta ORD, as detected by HA10 nm & GFP15 nm gold antibodies. Insets show ORP1L labelling in the membrane contact site between ER & autophagosome. The membranes of the ER are depicted in the bottom inset. Scale bar, 50 nm. (c) Three-colour super-resolution image of an autophagosomal vesicle labelled by LC3 (green), ORP1L (blue) & the ER protein VAP-A (red). Scale bar, 500 nm (d) MelJuSo cells cultured either in lipid depleted serum or control medium were fixed & stained for LC3 & ER marker Calnexin. Scale bar, 10 μm. Right panel: Manders coefficient for LC3 localization to the ER was calculated on at least 10 cells over three independent experiments. Bars indicate mean+s.d. Student's t-test statistical analysis (****P<0.0001). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27283760), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: LC3B Antibody - BSA Free [NB100-2220] - OSI induces autophagy through upregulation of MCT1 in CRC cells.a Immunoblotting analysis of MCT1 expression in CRC cells treated with or without 5 μM OSI for 24 h. b Immunoblotting analysis of MCT1 & phosphorylated AMPK in tumor xenografts obtained from vehicle- or OSI-treated mice. (Each protein of interest from each group was electrophoretically transferred onto a PVDF membrane, incubated with indicated primary & secondary antibodies, & developed as a digital image.) c Immunohistochemical analysis of MCT1 expression in tumor xenografts. Scale bar, 50 μm. d Relative intensity of MCT1 staining in (c). e CRC cells were transfected with siScramble or siMCT1 for 24 h, followed by treatment with or without 5 μM OSI for another 24 h. The protein levels of LC3, phosphorylated AMPK, phosphorylated LKB1 & MCT1 were analyzed by immunoblotting. f CRC cells were treated as in (e), the endogenous LC3 puncta were analyzed by immunofluorescence. Scale bar, 10 μm. g The number of LC3 puncta in (f). h CRC cells were transfected with empty vector or Flag-MCT1 plasmid for 48 h, the protein levels of MCT1 & phosphorylated AMPK were analyzed by immunoblotting. i CRC cells were treated as in (h), the endogenous LC3 puncta were analyzed by immunofluorescence. Scale bar, 10 μm. j The number of LC3 puncta in (i). k Immunoblotting analysis of LC3, MCT1 & phosphorylated AMPK levels in CRC cells co-transfected with Flag-MCT1 & DN-AMPK plasmids for 48 h. l CRC cells were treated as in (k), the endogenous LC3 puncta were analyzed by immunofluorescence. Scale bar, 10 μm. m The number of LC3 puncta per cell in (l). Data are presented as mean SEM, Student’s t-test, & are representative of three independent experiments. *P < 0.05; ***P < 0.001 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31409796), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Abrogation of autophagy by siRNAs targeting autophagy-related genes in gastric cancer cells induced tumor-intrinsic PD-L1 expression. a Inhibition of autophagy by knockdown of ATG5 in AGS & NCI-n87 gastric cancer cells induced the expression of PD-L1 in the presence & absence of INF-gamma (200 U/mL) as shown by flow cytometry analysis at 48 h post-transfection. b The induction of PD-L1 was confirmed by Western blots at 72 h post-transfection. The knockdown efficacies of ATG5 & ATG7 siRNA were verified. The conversion of LC3B-I to LC3B-II was reduced. Results were averaged & blots were representative of 4 independent experiments. The ratio of PD-L1 MFI minus isotype control was shown as mean ± S.D. relative to Ctrl from 4 independent experiments, *p < 0.05 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30925913), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - OSI induces autophagy through upregulation of MCT1 in CRC cells.a Immunoblotting analysis of MCT1 expression in CRC cells treated with or without 5 μM OSI for 24 h. b Immunoblotting analysis of MCT1 & phosphorylated AMPK in tumor xenografts obtained from vehicle- or OSI-treated mice. (Each protein of interest from each group was electrophoretically transferred onto a PVDF membrane, incubated with indicated primary & secondary antibodies, & developed as a digital image.) c Immunohistochemical analysis of MCT1 expression in tumor xenografts. Scale bar, 50 μm. d Relative intensity of MCT1 staining in (c). e CRC cells were transfected with siScramble or siMCT1 for 24 h, followed by treatment with or without 5 μM OSI for another 24 h. The protein levels of LC3, phosphorylated AMPK, phosphorylated LKB1 & MCT1 were analyzed by immunoblotting. f CRC cells were treated as in (e), the endogenous LC3 puncta were analyzed by immunofluorescence. Scale bar, 10 μm. g The number of LC3 puncta in (f). h CRC cells were transfected with empty vector or Flag-MCT1 plasmid for 48 h, the protein levels of MCT1 & phosphorylated AMPK were analyzed by immunoblotting. i CRC cells were treated as in (h), the endogenous LC3 puncta were analyzed by immunofluorescence. Scale bar, 10 μm. j The number of LC3 puncta in (i). k Immunoblotting analysis of LC3, MCT1 & phosphorylated AMPK levels in CRC cells co-transfected with Flag-MCT1 & DN-AMPK plasmids for 48 h. l CRC cells were treated as in (k), the endogenous LC3 puncta were analyzed by immunofluorescence. Scale bar, 10 μm. m The number of LC3 puncta per cell in (l). Data are presented as mean SEM, Student’s t-test, & are representative of three independent experiments. *P < 0.05; ***P < 0.001 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31409796), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - Screening of potent NAPIs in HepG2. a Immunoblots for autophagy-related proteins LC3-II, p62 (left); semi-quantified analysis (n = 3) in various nanoparticles treated cells (right). CQ & Rapamycin (Rapa) were used as positive controls for autophagy inhibition & autophagy activation, respectively. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control. Normalized band densities were shown below each band. b Fluorescence images of mCherry-GFP-LC3 cells after incubation with CQ or NDs for 48 h (autophagosomes: mCherry+/GFP+ yellow puncta; autolysosomes: mCherry+/GFP) & quantification of the number of LC3 puncta per cell in cells (10 cells per group). Scale bar: 10 μm. c Immunoblots for autophagy-related proteins LC3-II, p62 (left); semi-quantified analysis (n = 3) in CQ, NDs, or CQ–NDs-treated cells (right). &P < 0.05, significantly different from NDs. GAPDH was used as the loading control. Normalized band densities were shown below each band. d Left: Cell viability after incubation with ATO or various NAPIs–ATO mixture for 48 h (n = 3). ##P < 0.01 by t-test, significantly different from ATO. Right: Cell viability after 48 h NDs–ATO treatment with RNAi of autophagy proteins ATG5 & ATG7 (n = 3). e, f Immunoblots for autophagy-related protein LC3-II & autolysosomal process-related protein NUPR1, SNAP25, VAMP8 in NDs-treated cells. g Immunoblots for autolysosomal process-related protein NUPR1 after NDs treatment with RNAi of autophagy proteins ATG5 & ATG7. GAPDH was used as the loading control. Error bars are s.d. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30341298), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Screening of potent NAPIs in HepG2. a Immunoblots for autophagy-related proteins LC3-II, p62 (left); semi-quantified analysis (n = 3) in various nanoparticles treated cells (right). CQ & Rapamycin (Rapa) were used as positive controls for autophagy inhibition & autophagy activation, respectively. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control. Normalized band densities were shown below each band. b Fluorescence images of mCherry-GFP-LC3 cells after incubation with CQ or NDs for 48 h (autophagosomes: mCherry+/GFP+ yellow puncta; autolysosomes: mCherry+/GFP) & quantification of the number of LC3 puncta per cell in cells (10 cells per group). Scale bar: 10 μm. c Immunoblots for autophagy-related proteins LC3-II, p62 (left); semi-quantified analysis (n = 3) in CQ, NDs, or CQ–NDs-treated cells (right). &P < 0.05, significantly different from NDs. GAPDH was used as the loading control. Normalized band densities were shown below each band. d Left: Cell viability after incubation with ATO or various NAPIs–ATO mixture for 48 h (n = 3). ##P < 0.01 by t-test, significantly different from ATO. Right: Cell viability after 48 h NDs–ATO treatment with RNAi of autophagy proteins ATG5 & ATG7 (n = 3). e, f Immunoblots for autophagy-related protein LC3-II & autolysosomal process-related protein NUPR1, SNAP25, VAMP8 in NDs-treated cells. g Immunoblots for autolysosomal process-related protein NUPR1 after NDs treatment with RNAi of autophagy proteins ATG5 & ATG7. GAPDH was used as the loading control. Error bars are s.d. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30341298), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - OSI induces autophagy in CRC cells in vitro & in vivo.a Immunoblotting analysis of LC3, Atg5, & p62/SQSTM1 expression in CRC cells treated with indicated concentrations of OSI for 24 h. b The formation of endogenous LC3 puncta in cells treated with DMSO or 5 μM OSI for 24 h. c Total number of endogenous LC3 puncta per cell in (b). d, e LC3 expression in xenograft tissues was examined by IHC. Representative images were provided as indicated in (d) & relative intensity of LC3 staining was quantified in (e). f Immunoblotting analysis of LC3 & p62/SQSTM1 expression in tumor xenografts (Each protein of interest from each group was electrophoretically transferred onto a PVDF membrane, incubated with indicated primary & secondary antibodies, & developed as a digital image.) g Relative intensity of LC3 in (f). h Co-immunoprecipitation analysis of the interaction between Beclin 1 & Bcl-2 in CRC cells treated with or without 5 μM OSI for 24 h. i Immunoblotting analysis of LC3 expression in CRC cells treated with or without 5 μM OSI in the presence or absence of 5 mM 3-MA for 24 h. j CRC cells were treated as in (i), the LC3 puncta were analyzed by immunofluorescence. Scale bar, 10 μm. k, l Immunoblotting analysis of LC3 expression in CRC cells transfected with siScramble, siATG5 (k), or siBECN1 (l) for 24 h, followed by treatment with or without 5 μM OSI for another 24 h. m CRC cells were treated as in (k, l). The LC3 puncta were analyzed by immunofluorescence. Scale bar, 10 μm. Data are presented as mean SEM, Student’s t-test, & are representative of 3 independent experiments. **P < 0.01; ***P < 0.001 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31409796), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: LC3B Antibody - BSA Free [NB100-2220] - OSI induces autophagy in CRC cells in vitro & in vivo.a Immunoblotting analysis of LC3, Atg5, & p62/SQSTM1 expression in CRC cells treated with indicated concentrations of OSI for 24 h. b The formation of endogenous LC3 puncta in cells treated with DMSO or 5 μM OSI for 24 h. c Total number of endogenous LC3 puncta per cell in (b). d, e LC3 expression in xenograft tissues was examined by IHC. Representative images were provided as indicated in (d) & relative intensity of LC3 staining was quantified in (e). f Immunoblotting analysis of LC3 & p62/SQSTM1 expression in tumor xenografts (Each protein of interest from each group was electrophoretically transferred onto a PVDF membrane, incubated with indicated primary & secondary antibodies, & developed as a digital image.) g Relative intensity of LC3 in (f). h Co-immunoprecipitation analysis of the interaction between Beclin 1 & Bcl-2 in CRC cells treated with or without 5 μM OSI for 24 h. i Immunoblotting analysis of LC3 expression in CRC cells treated with or without 5 μM OSI in the presence or absence of 5 mM 3-MA for 24 h. j CRC cells were treated as in (i), the LC3 puncta were analyzed by immunofluorescence. Scale bar, 10 μm. k, l Immunoblotting analysis of LC3 expression in CRC cells transfected with siScramble, siATG5 (k), or siBECN1 (l) for 24 h, followed by treatment with or without 5 μM OSI for another 24 h. m CRC cells were treated as in (k, l). The LC3 puncta were analyzed by immunofluorescence. Scale bar, 10 μm. Data are presented as mean SEM, Student’s t-test, & are representative of 3 independent experiments. **P < 0.01; ***P < 0.001 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31409796), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Growth retardation & shortened lifespan of the mice with deletion of an autophagy-essential gene Atg7 in hematopoietic system. (A) Three genotypes for wild-type, heterozygote, & homozygote for Atg 7 deletion in hematopoietic system with representative images of the mice. The images were taken at age of 10 weeks. (B) PCR Genotyping analysis of the offsprings from Atg7f/f mice crossing Vav-iCre mice to screen Atg7f/f;Vav-iCre mice. The sequences for the primers used in PCR are given in the method section, & their PCR amplified bands representing specific genotypes were indicated in the agarose gel electrophoresis films. (C) Examination of Atg7 expression in wild-type & the Atg7-deleted mice. Upper panel, quantitative PCR analysis of Atg7 transcription normalized to Gapdh transcript in different organs; lower panel, western blotting analysis of autophagy-essential protein ATG7 & lipidation of LC3 in different organs. GAPDH used as a loading control. (D) Growth comparison between wild-type & Atg7-deleted mice. Wild-type mice progressively gain weight before age of 60 weeks (left panel), but Atg7-deleted mice cease weight gain at about age of 6 weeks (right panel). (E) Measurement of lifespan of wild-type & Atg7-deleted mice. (F) Immunohistological examination of heart, liver, lung & thymus from 10 weeks old wild-type & Atg7-deleted mice by HE staining. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31327762), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Overexpression of wild-type MAGEA6, but not mutant MAGEA6, suppresses autophagy in PDAC cell lines.(A) Immunoblot analysis of autophagy signaling in HPDE-iKRAS cells expressing GFP (Vec) & MAGEA6 variants. (B) Immunofluorescence staining of LC3B puncta in the transduced HPDE-iKRAS cells. Representative photos (left) & statistical analysis (mean ± standard deviation of counted cells, N=~100 per cohort) are shown. *p=0.002; two-tailed unpaired t-test. (C) Immunoblot analysis of autophagy substrate SQSTM1/p62 in the transduced HPDE-iKRAS cells treated with BafA1 for the indicated time points., Immunoblot analysis of (D) autophagy signaling & (E) SQSTM1/p62 accumulation in wild-type MAGEA6 expressing & (F) autophagy signaling in MAGEA6H305fs* expressing HPDE-iKRAS cells under nutrient-deficient conditions. (G) Immunoblot analysis of autophagy signaling in wild-type MAGEA6 expressing cells under prolonged nutrient-deficient conditions.Immunoblot analysis of autophagy signaling of transduced AsPC-1 & MIA PaCa-2.Immunoblot analysis of the accumulation of autophagy substrate SQSTM/p62 in the transduced HPDE-iKRAS cells under BafA1 for the indicated time points.Immunoblot analysis of autophagy activity in transduced HPDE-iKRAS cells under nutrient-depleted conditions as indicated. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32270762), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Inhibition of LRRK2 alters the autophagy/lysosomal pathway in H4 neuroglioma cells. A) LC3-II levels increase upon LRRK2-in1 inhibitor treatment (1 μM, overnight treatment; quantification from 3 independent replicates, the plot shows mean & SD, * indicates significance (p < 0.05)). B) Dose dependent increase in LC3-II upon overnight treatment with LRRK2-in1. C) MTT assay showing no alteration of cell viability upon over night treatment with LRRK2-in1 from 1 to 5 μM. A small toxic effect appeared with the higher dose (10 μM). The plot shows mean & SD, * indicates significance (p < 0.05). D) LRRK2 knockdown cells display reduced response to LRRK2-in1. LRRK2 protein levels are decreased in shRNA stable line compared to wild type cells (right panel), & knockdown of LRRK2 reduces response to 1 μM LRRK2-in1 treatment compared to wild type or scrambled shRNA cells (left panel). E) Western blot analysis of H4 cells treated with DMSO & LRRK2-in1 (5 μM, 2.5 hours treatment) in the presence & absence of 40 nM bafilomycin added at the same time as the inhibitor. Quantification of three replicates is shown in the right hand panel, the plot shows mean & SD, * indicates significance (p < 0.05). F) LRRK2-in1 increases LC3-II levels independent of mTORC1 activity. P70S6K & phoshoThr389-P70S6K; S6 & phosphoSer235/236-S6 levels are shown in control, starvation & amino-acid stimulated conditions. LRRK2-in1 (1 μM over night) treatment does not alter phosphorylation of P70S6K & S6 in conditions that show increased levels of LC3-II. G) Neutral red staining. The plot shows mean & SEM. * indicates significance (p < 0.05). Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S0167488913002954), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - SQSTM1 recruited ARHGAP5-AS1 for autophagic degradation.a ARHGAP5-AS1 expression in resistant cells after transfecting siNC or SQSTM1 siRNAs was assessed using qRT-PCR. **p < 0.01. b ARHGAP5-AS1 expression in sensitive cells after SQSTM1 overexpression was detected by qRT-PCR. c The interaction of ARHGAP5-AS1 with SQSTM1 in SGC-R cells was analyzed by RIP assay followed with qRT-PCR. IgG was served as the negative control. d RNA pull down assay was performed to verify the binding of SQSTM1 to ARHGAP5-AS1 in SGC7901 or SGC-R cells. beta -actin was served as negative control. NC: no probe. The interaction of ARHGAP5-AS1 with SQSTM1 in SGC-R cells before & after EBSS treatment (4 h) were analyzed using RIP assay (e) & RNA pull down assay (f). g The interaction of various SQSTM1 constructs with ARHGAP5-AS1 were analyzed using RIP assay. h The interaction of various ARHGAP5-AS1 fragments with SQSTM1 was analyzed by RNA pull down assay. i Half-life of ARHGAP5-AS1 in SGC7901 & SGC-R cells with different SQSTM1 expression status were determined by qRT-PCR. The colocalization of ARHGAP5-AS with SQSTM1 (j) & LC3B (k) in SGC-R cells treated as indicated were analyzed by combined FISH & IFC assay (original magnification, ×100). Scale bar: 100 μm (j) & 10 μm (k) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31097692), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Effects of autophagy & caspase inhibitors on survival of neurons after glucose reperfusion (GR).After 3 h GD, neuro2a cells were replaced by 20-Methyladenine (3MA); Bafilomycin A1 (BA1) & z-VAD-FMK (z-VAD) were added at final concentrations of 5 mM, 5 nM & 20 uM, respectively. (A) Cell viability was assessed using the MTT assay after treatment of autophagy & pan-caspase inhibitors. MTT mitochondrial reduction was shown as a relative percentage of the MTT values at the indicated time point (3 h GD/48 h GR) from three independent experiments ± SD (*p<0.05, **p<0.001). (B) Changes in protein levels by inhibitors of autophagy or caspase activation were analyzed by immunoblot. Proteins were blotted with anti-LC3, p62, cleaved-caspase 3 & PARP antibodies. Tubulin was used as loading control. (C) Quantification of immunoblot data in (B). The amount of each protein was normalized against the amount of tubulin. Data represent the mean ± SD (*p<0.05, n = 3). (D) Cells transiently expressing GFP-LC3 (green) were treated with 3MA, BA1 or z-VAD for 48 h of GR after 3h GD. Nuclei were stained with DAPI (blue). Scale bar represents 5 µm. Quantification shown on the right graph represents mean GFP-LC3 puncta per cell (n = 10 cells per condition) from three independent experiments ± SD. The asterisks (*) indicate significant differences in the values.*p<0.05, **p<0.001. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0076466), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - Exposure of microglia to cART resulted in blockade of autophagosome–lysosome fusion. (A) rPMs were seeded into a 12-well plate followed by tandem fluorescent-tagged MAP1LC3B plasmid. Next, cells were exposed to cART (5 µM each of TDF, FTC, & DTG) for an additional 24 h & observed by confocal imaging. The results showed that cART exposure significantly increased the formation of autophagosomes (yellow puncta). (B) Representative bar graph showing the number of autophagosome (yellow puncta) per cell. (C) Representative bar graph showing the number of autolysosome (red puncta) per cell. (D) rPMs were seeded into 12-well plates followed with cART exposure for 24 h. Cells were then double immunostained with MAP1LC3B & LAMP2 antibody & observed by immunofluorescent microscopy. (E,F) Representative bar graphs showing cART-mediated decreased LAMP2 puncta & decreased colocalization of MAP1LC3B & LAMP2. BAF—autophagosome fusion inhibitor, & rapamycin (RAP—autophagy inducer) were used as controls for autophagy flux. Data is from three independent experiments & is expressed as means ± SEM & were analyzed using one-way ANOVA. *, p < 0.05 vs. control. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31569373), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - The cytoprotection of in vitro sIPC is diminished by autophagy inhibitors in RPTC cells. RPTC cells were subjected to: (1) control; (2) CCCP-R; (3) sIPC + CCCP-R in the absence or presence of chloroquine (20 μM) & 3-methyladenine (10 mM). Both inhibitors were used for 1-h pretreatment & during 2-h recovery from prolonged CCCP treatment. Cells were collected for morphological & immunoblot analyses. (a) Representative images of phase contrast & fluorescence microscopy showing cellular & nuclear morphology of apoptosis. Scale bar: 200 μm. (b) Quantification of cell apoptosis. Data are expressed as mean ± SD. *, P < 0.05, significantly different from the control group; #, P < 0.05, significantly different from CCCP-R group. (c) Analysis of apoptosis inhibitory efficiency by sIPC. Data are expressed as mean ± SD. *, P < 0.05, significantly different from the group without inhibitors. (d & e) Immunoblots of LC3B & cleaved CASP3. ACTB was used as a loading control. The molecular mass marker lanes were labelled as kDa. For densitometric analysis of cleaved CASP3, after normalization with ACTB, the protein signals of the control were arbitrarily set as 1, & the signals of other conditions were normalized to the control to calculate fold changes. Data are expressed as mean ± SD. *, P < 0.05, significantly different from the control group; #, P < 0.05, significantly different from CCCP-R group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31066324), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - TFEB overexpression in macrophages induces the autophagy markers LC3 & p62 & restores their co-localization in atherosclerotic aortic roots.(a,b) Representative immunofluorescence images of atherosclerotic aortic roots (2 months' western diet) from control & mφTFEB-TG mice (ApoE-null background) stained with antibodies against TFEB (a), TFEB & MOMA-2 (b; scale bar, 50 μm). (c) Quantification of the average TFEB intensity & co-localization with nuclear marker DAPI (n=4-5 mice per group). (d) Representative immunofluorescence images of atherosclerotic aortic roots from control & mφTFEB-TG mice stained with p62 & LC3 (scale bar, 50 μm). (e) Quantification of the p62 & LC3 average intensity from control & mφTFEB-TG-stained roots (n=13–14 mice per group). (f) Representative pseudocolour image of these p62/LC3 images (green represents co-localization) & graph depicting the increased p62/LC3 correlation seen in a representative mφTFEB-TG as compared to a control lesion (scale bar, 50 μm). (g) Quantification of the p62/LC3 co-localization from control & mφTFEB-TG-stained roots shown (n=13–14 mice per group). (h,i) FACS analysis of aortic macrophages isolated from atherosclerotic aortas of Control or mφTFEB-TG mice (western diet-fed ApoE-KO background, n=3–4 pooled aortas) & stained for either (h) p62 & LC3, or (i) Lamp2 & LC3 antibodies (per cent of macrophages expressing each marker is shown below plots). For all graphs, data are presented as mean±s.e.m. *P<0.05, ***P<0.001, two-tailed unpaired t-test. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms15750), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: LC3B Antibody - BSA Free [NB100-2220] - Keap1 co-localizes with SQSTM1-immunopositive sarcoplasmic aggregates in AVM muscle. a-c. Only focal & minimal co-localization of LC3 (a) & Keap1 immunofluorescence (b) is seen in a representative toxic AVM sample (colchicine-treated subject #31); merged panel is shown in (c) (LC3, red; Keap1, green). d-f. In the same specimen, there is extensive co-localization of SQSTM1 (d) & Keap1 immunofluorescence (e); merged panel is shown in (f) (SQSTM1, red; Keap1, green). g-i. Similar to toxic AVM specimens, essentially no co-localization of LC3 (g) & Keap1 immunofluorescence (h) is seen in a representative IBM muscle biopsy (subject #46); merged panel is shown in (i) (LC3, red; Keap1, green). j-l. In the same specimen, there is extensive co-localization of SQSTM1 (j) & Keap1 immunofluorescence (k); merged panel is shown in (l) (SQSTM1, red; Keap1, green). Scale bar, 25 μm Image collected & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/s40478-016-0384-6), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Glucose deprivation (GD) induces neuronal cell death.Time course of cell viability & cell death after GD was estimated by MTT & LDH assays, respectively. Bars depict rates of MTT reduction (A) & LDH release (B) time dependent manner after GD. MTT data represent relative percentages compared with controls (*p<0.05, **p<0.001, n = 3). LDH release from neuro2a cells was calculated as fold increase from the LDH release in controls (*p<0.01, **p<0.001, n = 3). (C) Levels of LC3 & p62 expression, cleaved-caspase 3 & PARP cleavage were determined at indicated times after GD by immunoblot analysis. Tubulin was used as loading control. (D) Quantitative analyses of the immunoblots are shown in (C). The amount of each protein was normalized against the amount of tubulin. Data represent the mean ± SD for each condition (*p<0.05, **p<0.001, n = 3). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0076466), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - Measurement of dysregulated autophagy by fluorescent microscopyOVCAR3 cells were treated with the indicated drugs for 24 hours & then fixed & immunostained for p62 (red) & LC3 (green), with a DAPI costain (blue). Confocal z-stacks were flattened for image analysis to capture all punctate area. A. Doses were: chloroquine (10 μM), nelfinavir (10 μM), rapamycin (10 nM), dasatinib (50 nM), & all were combined for Necramed. B. Similar immunostaining as in (A) Punctae were quantified for size & number per cell by ImageJ with at least 40 cells per condition. LC3 channel was analyzed for autophagosomes, & the p62 channel for sequestosomes. Doses were same as in (A), but with chloroquine & nelfinavir reduced to 5 μM. *p < 0.05, ***p < 0.001 by t-test. All error bars are s.e.m. C. OVCAR3 cells with a virally integrated mCherry-GFP-LC3 construct were studied by live microscopy, for the number of hours indicated following Necramed (doses as in (A)) or control treatment. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26418751), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Rop-DPRL combined with CR inhibits tumor growth. (A) Fluorescent imaging of major organs & tumor. Tumor-bearing mice were injected with cy5-labeled liposome or unlabelled liposome. The tumor & major organs from control or drug-treated mice were excised at 12 h or 24 h. (B) Fluorescent image of tumor sections. Tumor from control or Rop-DPRL treated mice were excised at 12 h. Tumor frozen sections were observed under confocal microscope. Scale bar = 20 µm. (C) Immunofluorescence staining of tumor sections for the autophagosome marker LC3, P62 & LAMP1. (D) Image of representative tumors. (E) Change in tumor volume. (F) Tumor weight. (G) TUNEL staining of tumor sections. Nuclei were stained with Hoechst. Scale bar = 20µm. (H) Body weight changes of mice under different treatments. CR: Calorie restriction. Arrows: Enlarged lysosomes. Data are presented as the mean ± SEM, ** p < 0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32308756), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - SsD & SsA inhibit autophagy in HeLa cells. a Structures of SsA, SsC, & SsD. b SsD (15 μM) & SsA (30 μM), but not SsC, significantly increased LC3-II yellow puncta formation but did not significantly affect LC3-II red-only puncta in RFP-GFP-LC3-expressing HeLa cells. Scale bar = 10 μm. Quantification of LC3 yellow puncta/red puncta (%) is presented as the mean ± S.E., n = ~80 cells from 3 independent experiments. c Treatment of HeLa cells with SsD for 6 h induced the accumulation of both LC3-II & SQSTM1 in a dose-dependent manner. d SsD (15 μM) induced the accumulation of both LC3-II & SQSTM1 in HeLa cells in a time-dependent manner. e SsA (30 μM) induced the accumulation of both LC3-II & SQSTM1 in HeLa cells in a time-dependent manner. f Treatment with different concentrations of SsC for 6 h failed to induce the accumulation of either LC3-II or SQSTM1 in HeLa cells. g SsD (15 μM) reversibly inhibited autophagy Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30820356), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - PERK pathway-independent induction of prosurvival autophagy in KG1 LSC-like cells upon G9a inhibition. a, LC3-I/II & p62 expression, analyzed by western blotting, after cell treatment with BIX-01294 (15 μmol/L) in the absence or presence of bafilomycin A1 (BafA1) for 24 h. b, Confocal microscopy images of GFP-LC3-transfected cells after treatment with BIX-01294 (5 μmol/L). c, Apoptotic fractions, measured using flow cytometry, after cell treatment with BIX-01294 (15 μmol/L) in the absence or presence of the autophagy inhibitor 3-MA (5 mmol/L) or BafA1 (2 nmol/L) for 48 h. d, Protein expression, analyzed by western blotting with the indicated antibodies, after treatment of PERK siRNA-transfected & non-transfected cells with BIX-01294 (15 μmol/L) for 48 h. e, Protein expression, analyzed by western blotting with the indicated antibodies, after cell treatment with BIX-01294 (10 μmol/L) in the absence or presence of GSK2606414 (20 μmol/L) for 48 h. f, Protein expression, analyzed by western blotting with the indicated antibodies, after treatment of NRF2 siRNA-transfected & non-transfected cells with BIX-01294 (10 μmol/L) for 48 h. g, Levels of apoptosis, evaluated by flow cytometry, after cell treatment for 48 h with BIX-01294 (10 μmol/L) in the absence or presence of the PERK inhibitor GSK2606414 (10 μmol/L) or the autophagy inhibitor 3-MA (5 mmol/L), or BafA1 (2 nmol/L). h, Levels of apoptosis, evaluated by flow cytometry, after treatment of PERK siRNA- or control siRNA-transfected cells with BIX-01294 (10 μmol/L) in the absence or presence of 3-MA (5 mmol/L) or BafA1 (2 nmol/L) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32293500), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Metabolic profiling & biochemical assay. (A) Relative abundance of the substrates in the glycolytic pathway & TCA cycle in ORP compared to OCL by targeted metabolic profiling. When compared with OCL heart, ORP hearts have significantly lower glucose-6-phosphate & fructose-6-phosphate (both are glycolytic metabolites), & significantly higher a-ketoglutarate, fumarate, malate, & citrate (all are TCA cycle metabolites). *P < 0.05 compared with OCL. See Table S6 for numerical data. (B) A schematic diagram summarizing the changes in metabolism by rapamycin in old heart. (C) Western blots of autophagic markers show no significant change of LC3 II/I, p62, or beclin-1 in cardiac aging. However, OCR has significantly lower p62 than that in OCL. #P < 0.05 compared with OCL. (D) Both CR & RP significantly reduce the age-dependent increase in protein carbonyls (nmol mL−1). #P < 0.05 compared with OCL. (E). Both CR & RP significantly reduce the age-dependent increase in protein ubiquitination.*P < 0.05 compared with YCL & #P < 0.05 compared with OCL. n = 3–8. G6P: glucose 6-phosphate; G1P: glucose 1-phosphate; F6P: fructose 6-phosphate; F1P: fructose 1-phosphate; F16BP: fructose 1,6-bisphosphate; F26BP: fructose 2,6-biphosphate; G3P: glyceraldehyde 3-phosphate; DHAP:dihydroxyacetone phosphate; 2(3)-PGA: 2- or 3-phosphoglycerate; & PEP: phosphoenolpyruvate. Isomers of same molecular weight, that is, G6P versus G1P, F6P versus F1P, & F16BP versus F26BP, were not distinguishable by the LC-MS/MS-based metabolic profiling method. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24612461), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - iNOS level is regulated by autophagy during LPS-stimulated condition. (A) Raw 264.7 cells were treated with LPS (200 ng/mL) for 12 h first, followed by removing the LPS & added CQ (30 µM), SAR (1 µM), Torin (1 µM), Rap (1 µM) for another 12 h. CTRL is blank control group with DMSO treatment. (B) Western blot analysis of iNOS in Raw 264.7 cells under LPS treatment. (C) Western blot analysis of p62 in Raw 264.7 cells under LPS treatment. (D) Raw 264.7 cells were treated LPS (200 ng/mL) for 12 h, then removed LPS added CHX (1 µg/mL) & autophagy related drugs for another 12 h. (E) Level of iNOS in Raw 264.7 cell after CHX treatment. (B–E) Values were analyzed from three individual experiments by GraphPad Prism, each experiment conducted three times. * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars (mean ± SEM). One-way ANOVA with Turkeys as post hoc tests. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31618870), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - Autophagic pathway regulates inflammasome activation in blood-derived monocytes stimulated with M. leprae. (A–C) Monocytes were purified from healthy donors’ peripheral blood mononuclear cells by 2-h adherence & stimulated with 10 µg/mL M. leprae (ML) in the presence or absence of autophagy regulators for 18 h. Autophagy was triggered with 200 ng/mL rapamycin (RP), & inhibited with 100 µM chloroquine (CQ, autophagic flux blocker) & 10 µM 3-methyladenine (3-MA). (A) Monocytes stimulated with M. leprae, RP, CQ, & 3-MA were fixed & stained with anti-NLRP3 (red), anti-LC3B (green) antibodies & DAPI (blue). Monocytes stimulated with M. leprae increase the number of LC3-II-decorated autophagosomes (puncta) per cell, which is reforced by RP addition, & reverted by 3-MA. Autophagy blocking with CQ & 3-MA was able to increase NLRP3 specks numbers per cell in relation to the well stimulated with M. leprae & RP. Immunofluorescence images were quantified & bars represent the mean values of the number of LC3 puncta & NLRP3 specks per cell ± SEM (n = 3). *(in relation to NS) or #(indicated by the dashes) p < 0.05. Scale bar: 10 µm. (B) Purified mRNAs from monocyte cultures stimulated with M. leprae, the autophagy blocker 3-MA, & RP, as a control of autophagy activation, were analyzed by RT-qPCR for LC3B & IL1B. Bars represent the mean of the fold change values in relation to the non-stimulated controls of three independent experiments with similar results. (C) The IL-1 beta , IL-6, & TNF levels in the monocyte cultures supernatants were assessed by ELISA. Bars represent the fold change values in relation to the non-stimulated controls ± SEM (n = 3). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29915584), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - Melatonin induced LC3 aggregation & conversion. a GFP-LC3 aggregation & conversion in U87MG cells stably expressing an exogenous fusion protein, green fluorescent protein (GFP) & microtubule-associated light chain 3 (LC3) (U87-GFP-LC3), after treatment with DMSO (control), rapamycin (RAP 200 nM), melatonin (MEL 1 mM) or agomelatine (AGO 2 μM) for 24 h (×40); b U87MG cells & c A172 cells treated with DMSO (control), rapamycin (RAP 200 nM), melatonin (MEL 1 mM), agomelatine (AGO 2 μM) & luzindole (LUZ 5 μM/L) for 24 h were fixed & analyzed for endogenous LC3 dots by immunofluorescence using anti-LC3 antibodies (green), DAPI (blue) staining shows the nuclei (×40). d Stable U87-GFP-LC3 cell lines & GFP in vector control. U87MG cells stably expressing an exogenous fusion protein, green fluorescent protein (GFP) & microtubule-associated light chain 3 (LC3) (U87-GFP-LC3) (×40) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31870319), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - Autophagy induction by atorvastatin in T24 human bladder cancer cells. (A) Immunocytochemistry for the co-localization of LC3 puncta (green) & Lysotracker (red, demarcation for the lysosome) after atorvastatin treatments (30 μM) for 24 h in T24 cells. Rapamycin treated T24 cells were the positive control of autophagy induction. 4′,6-diamidino-2-phenylindole (DAPI) was used for nucleus staining. Scale bar = 100 μm; (B) High magnification view of immunocytochemistry for the co-localization LC3 puncta (green) & Lysotracker after atorvastatin treatments (30 μM) for 24 h in T24 cells at the single-cell level. Scale bar = 50 μm; (C) Western blot analysis of autophagosome formation markers p62/SQSTM1, LC3-I & LC3-II in untreated (control) & atorvastatin (30 μM) treated T24 cells. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/15/5/8106), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - Increased LC3B-positive puncta correlated positively with vascular remodeling during AngII-induced hypertension. (A) Western blot analysis of LC3B-II expression in aortas from sham or AngII-infused mice at different time points after operation. **P < 0.01 vs. the corresponding sham group, Student's t-test. n = 10 mice/group. (B) Representative immunofluorescence staining of LC3B (green) & alpha -SMA (red) in thoracic aorta section. Nuclei were stained with DAPI. Scale bars, 20 µm. (C) Quantification of LC3B puncta. Five random fields (2,500 µm2/filed) were measured in one section. Statistical significance was determined by one-way ANOVA. **P < 0.01 vs. sham, n = 12 sections from six mice per group. (D-G) Structural parameters of mouse aorta from sham & hypertensive mice. WD, wall diameter; LD, lumen diameter; CSA, cross-sectional area. *P < 0.05, **P < 0.01 vs. the corresponding sham group, Student's t-test. n = 10 mice/group. (H) Correlation between changes in LC3B puncta & CSA values. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32226533), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: LC3B Antibody - BSA Free [NB100-2220] - Representative images of the variation of LC3 staining associated with cavities in SDH activity & mitochondrial COX-1 staining in PAD gastrocnemius muscle sections. First column shows succinate dehydrogenase (SDH) activity (dark fibers are type I, intermediate fibers are type IIa, & light fibers are type IIa/x), second column shows mitochondrial COX-1 protein (green), third column shows LC3 (red) & fourth column shows COX-1 & LC3 merged. Examples of normal fibers with no LC3 staining or COX-1 cavities (rowA), punctate LC3 staining where a COX-1 cavitiy is forming (row B), elevated LC3 staining with low COX-1 staining (rowC), LC3 accumulation in areas lacking COX-1 staining (row D), & LC3 plaque in the center of an SDH/COX1 cavity (COX-1; rowE). Arrows within the same row point to areas of LC3 accumulation in same fiber. Scale bar = 100 µM Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27687713), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Mechanistic analysis of autophagic inhibition in ATO-treated HepG2. a The viability of HepG2 & NB4 cells after incubation with ATO at various concentrations for 48 h (n = 3; error bars are s.d.). Upper: Confocal images of NB4 or HepG2 cells after being stained with calcein AM. Scale bars: 10 μm. b The viability of HepG2 & NB4 cells after incubation with CQ, ATO, or CQ–ATO mixture for 48 h (n = 3). ##P < 0.01 by t-test, significantly different from ATO. ns not significant. c Immunoblots of autophagy-related proteins LC3-II, p62 (left); semi-quantified analysis (n = 3) in NDs, ATO, or NDs–ATO mixture-treated cells (right). GAPDH was used as the loading control. Normalized band densities were shown below each band. d Cell viability & Caspase-3 activity after incubation with ATO or NDs–ATO mixture for 48 h (n = 3). ##P < 0.01 by t-test, significantly different from ATO. e Immunostaining of LC3 in NDs, ATO, or NDs–ATO mixture-treated cells (left). Scale bar: 20 μm. TEM images of ATO or NDs–ATO mixture-treated cells (right). Scale bars: 1 μm. Typical structures of autophagosomes induced by ATO & autolysosome containing NDs induced by NDs–ATO mixture are indicated with green or blue arrows, respectively (magnification). Vacuoles inside cells are indicated with red arrows (magnification). Scale bars: 500 nm. Error bars are s.d. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30341298), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - Rop-DPRL impairs autophagy. (A) Fluorescent images of GFP-LC3-HeLa cells treated with 1.5 mM ropivacaine for 0-24 h, or starvation for 0-24 h, & stained with 75 nM LysoTracker Red. Scale bar = 5 µm. (B) Western blot & (C) statistical results of P62, immature & mature cathepsin D (CathD), LC3I, LC3II, & beta -actin. HeLa cells were treated with ropivacaine or starvation for 0-24 h. (D) Western blot & (E) statistical results of P62, LC3I, LC3II, & beta -actin. HeLa cells were treated with PBS, CQ (50 µM), ropivacaine, or CQ+Rop for 24 h. (F) Fluorescent images of GFP-LC3-HeLa cells treated with empty liposome or Rop-DPRL for 24 h & stained with 75 nM LysoTracker Red. Scale bar = 5 μm. (G,I, K & M) Western blot & (H,J,L & N) statistical results for P62, LC3I, LC3II & beta -actin. Cells were treated with empty liposome, CQ, Rop-DPRL, starvation or indicated combination for 24 h. Data are presented as the mean ± SEM, *p < 0.05, ** p < 0.01, *** p < 0.001, ns = no significant difference, Rop: ropivacaine, starv: starvation. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32308756), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: LC3B Antibody - BSA Free [NB100-2220] - PERK pathway-independent induction of prosurvival autophagy in KG1 LSC-like cells upon G9a inhibition. a, LC3-I/II & p62 expression, analyzed by western blotting, after cell treatment with BIX-01294 (15 μmol/L) in the absence or presence of bafilomycin A1 (BafA1) for 24 h. b, Confocal microscopy images of GFP-LC3-transfected cells after treatment with BIX-01294 (5 μmol/L). c, Apoptotic fractions, measured using flow cytometry, after cell treatment with BIX-01294 (15 μmol/L) in the absence or presence of the autophagy inhibitor 3-MA (5 mmol/L) or BafA1 (2 nmol/L) for 48 h. d, Protein expression, analyzed by western blotting with the indicated antibodies, after treatment of PERK siRNA-transfected & non-transfected cells with BIX-01294 (15 μmol/L) for 48 h. e, Protein expression, analyzed by western blotting with the indicated antibodies, after cell treatment with BIX-01294 (10 μmol/L) in the absence or presence of GSK2606414 (20 μmol/L) for 48 h. f, Protein expression, analyzed by western blotting with the indicated antibodies, after treatment of NRF2 siRNA-transfected & non-transfected cells with BIX-01294 (10 μmol/L) for 48 h. g, Levels of apoptosis, evaluated by flow cytometry, after cell treatment for 48 h with BIX-01294 (10 μmol/L) in the absence or presence of the PERK inhibitor GSK2606414 (10 μmol/L) or the autophagy inhibitor 3-MA (5 mmol/L), or BafA1 (2 nmol/L). h, Levels of apoptosis, evaluated by flow cytometry, after treatment of PERK siRNA- or control siRNA-transfected cells with BIX-01294 (10 μmol/L) in the absence or presence of 3-MA (5 mmol/L) or BafA1 (2 nmol/L) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32293500), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - TLR4 mediates Hsp70/Hsp90-induced catabolic response in myotubes. a Hsp70 & Hsp90-induced catabolic response in myotubes is dependent on TLR4. C2C12 myoblasts were transfected with siRNA as indicated. After differentiation, myotubes were treated with rHsp70 & rHsp90 & analyzed for catabolic response as described above. b LLC-released EVs induce muscle catabolism in C2C12 myotubes through TLR4. C2C12 myotubes with either TLR2 or TLR4 knockdown were treated with EVs isolated from LCM (AchE 6 mU) & analyzed for catabolic response. c CD9-positive EVs activate TLR4 in reporter cells. EVs isolated from LCM were subjected to immunoprecipitation (IP) using an antibody against CD9 with pre-immune IgG as control. Resulting pellet & supernatant were used to treat TLR4 reporter cell line HEK-Blue hTLR4 for 24 h, & compared with original EVs. TLR4 activation was measured as SEAP activity. Data (n = 3) were analyzed by analysis of variance. * denotes a difference (P < 0.05) Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-017-00726-x), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB100-2220] - cIAP2 & XIAP regulate autophagosome fusion, but not cIAP1.a MEFs expressing mCherry-GFP-LC3b were transfected with siRNA against either cIAP1, cIAP2 or XIAP. Cells were analysed with the microscope & the number of mCherry+, & GFP + puncta/cell were calculated & the ratio of GFP + /mCherry + puncta is indicated. Shown are the means & the error bars represent the SEM of at least three independent experiments. Westerns show efficient knockdown of cIAP1 & XIAP expression. cIAP2 siRNA efficiency was determined by real time PCR as shown in the graph below the westerns. b Wild type & cIAP2−/− XIAP−/− dermal fibroblasts expressing mCherry-GFP-LC3b were treated analysed on the microscope & mCherry+, & GFP + puncta/cell were calculated. The ratio of GFP+/mCherry + puncta is indicated. c Wild type & cIAP2−/− XIAP−/− dermal fibroblasts were left in complete media (CM) or starved for 2 h in EBSS. Cells were lysed & proteins analysed by western blot for XIAP, LC3, & Actin. cIAP2−/− was confirmed by PCR due to lack of effective antibodies for mouse cIAP2 (see supplemental Fig. 2). d Immunofluoresence showing accumulation of LC3 in starved cIAP2−/− XIAP−/− dermal fibroblasts. Wild type & cIAP2−/− XIAP−/− dermal fibroblasts were incubated in complete media or starved in EBSS for 2 h, then fixed & stained for LC3 (green channel) & LAMP2 (red channel). Nuclei are stained blue with Hoechst. Shown in upper panels are overviews. Lower panels show zoomed in regions indicated in the upper panels Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29743550), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Enhanced mTORC2 signaling is sufficient to protect MSC from calcification. Flag-tagged rictor or green fluorescent protein (GFP) as a control were introduced into MSC by lentiviral transfer. Cells were cultured for 21 days under control conditions, with osteoblast induction medium (OM) & vehicle, or OM & 20 nM Rapa as indicated. (A) Western blot analyses of rictor & flag confirming overexpression of rictor due to gene transfer, downstream targets of the two mTOR complexes (mTORC1: pp70-S6KSer389; mTORC2: pAKTSer473), inhibition of autophagy (LC3B II, p62), cellular senescence (p16INK4a), & apoptosis-related proteins (Bcl-2: negative regulator of apoptosis; cleaved caspase 3 (cl casp 3): executioner caspase). Representative western blots from six independent experiments are shown. For densitometric quantification, band intensities were normalized to GAPDH as a loading control. Control (no OM) was set to 1. (B) Lactate dehydrogenase (LDH) activity in cell culture supernatants normalized to total protein concentrations was determined to quantify cell death. Control was set to 1. (C) Alkaline phosphatase (ALP) activity normalized to total protein concentrations was measured after 7 days of incubation. (D) Calcium deposition was quantified with the ortho-cresolphthalein method & normalized to total protein concentrations. Bars represent mean + SEM, *P < 0.05, **P < 0.01, ***P < 0.001. Numbers of independent experiments are indicated at the bottom of each graph. (E) Alizarin red S staining of calcium deposits. Representative experiment out of six, phase contrast microscopy, original magnification x200, scale bar = 100 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31882658), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB100-2220] - OSI induces autophagy by activating LKB1/AMPK signaling in CRC cells.a Immunoblotting analysis of AMPK, phosphorylated AMPK (Thr172) & LC3 levels in CRC cells treated with indicated concentration of OSI for 24 h. b Immunohistochemistry analysis of AMPK phosphorylation levels in xenograft tissues. Scale bar, 50 μm. c Relative intensity of phosphorylated AMPK staining in (b). d CRC cells were transfected with siScramble or siAMPK for 24 h & followed by treatment with or without 5 μM for another 24 h. LC3 & phosphorylated AMPK (Thr172) levels were detected by immunoblotting. e CRC cells were transfected with empty vector or DN-AMPK plasmid for 24 h, followed by treatment with or without 5 μM OSI for another 24 h. LC3 & AMPK phosphorylation levels were determined by immunoblotting. f CRC cells were treated as in (d), the endogenous LC3 puncta in CRC cells were assessed by immunofluorescence. Scale bar, 10 μm. g The number of LC3 puncta per cell in (f). h CRC cells were treated as in (e), the endogenous LC3 puncta in CRC cells were assessed by immunofluorescence, Scale bar, 10 μm. i The number of LC3 puncta per cell in (h). j Immunoblotting analysis of phosphorylated LKB1 & phosphorylated CaMKK beta levels in CRC cells treated with or without 5 μM OSI. Data are presented as mean SEM, Student’s t-test, & are representative of three independent experiments. ***P < 0.001 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31409796), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: LC3B Antibody - BSA Free [NB100-2220] - Cathepsin inhibitors promote neuronal survival after glucose reperfusion (GR).After 3 h GD, neuro2a cells were replaced by 20; pepstatin A (PA, 10 uM), leupeptin (Leu, 10 uM) or both (PA+Leu). (A) Cell viability was assessed using the MTT assay. MTT mitochondrial reduction was shown as a relative percentage of the MTT values at the indicated time point (3 h GD/48 h GR) from three independent experiments ± SD (*p<0.05, **p<0.001). (B) Changes of protein level by cathepsin inhibitors were analyzed by immunoblot analysis. Proteins from cell lysates were fractionated on SDS-PAGE; blots were probed with anti-LC3, anti-p62, anti-cleaved caspase-3, & anti-PARP antibodies. Tubulin was used as loading control. (C) The graphs represent relative band density of each protein shown in (B). The amount of each protein was normalized against the amount of tubulin. Data represent the mean ± SD (*p<0.05, n = 3). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0076466), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt, Po, Al, Av, Ba, Bv, Ca, Ch, ChHa, Gp, Ha, In, Pm, Pm, Rb, SyHa, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Al, Av, Ba, Bv, Ca, Ch, ChHa, Gp, Ha, In, Pm, Pm, Rb, SyHa, Ze

Applications:

, , , , , , , , , , , ,

Applications: , , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-2220AF350
Alexa Fluor 405	NB100-2220AF405
Alexa Fluor 488	NB100-2220AF488
Alexa Fluor 532	NB100-2220AF532
Alexa Fluor 594	NB100-2220AF594
Alexa Fluor 647	NB100-2220AF647
Alexa Fluor 700	NB100-2220AF700
Alexa Fluor 750	NB100-2220AF750
Allophycocyanin	NB100-2220APC
Biotin	NB100-2220B
CoraFluor 1	NB100-2220CL1
DyLight 350	NB100-2220UV
DyLight 405	NB100-2220V
DyLight 488	NB100-2220G
DyLight 550	NB100-2220R
DyLight 594	NB100-2220DL594
DyLight 650	NB100-2220C
DyLight 680	NB100-2220FR
DyLight 755	NB100-2220IR
FITC	NB100-2220F
HRP	NB100-2220H
Janelia Fluor 525	NB100-2220JF525
Janelia Fluor 549	NB100-2220JF549
Janelia Fluor 646	NB100-2220JF646
Janelia Fluor 669	NB100-2220JF669
PE	NB100-2220PE
PerCP	NB100-2220PCP
mFluor Violet 450 SE	NB100-2220MFV450
mFluor Violet 500 SE	NB100-2220MFV500
mFluor Violet 610 SE	NB100-2220MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-2220SS

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NB300-221

GAPDH Antibody (1D4) - BSA Free

(19 Reviews)

(455 Publications)

0.1 ml

NB300-221

Analysis of GAPDH in human nucleus pulposus cell culture (40ug/lane) using anti-GAPDH antibody. LI-COR Odyssey imaging system was used for detection. Western blot image submitted by a verified customer review.

Analysis of of HeLa cells stained with GAPDH antibody, dilution 1:100 (Green). DAPI staining of nuclear DNA (Blue). The GAPDH antibody produces strong cytoplasmic staining of healthy cells.

Analysis of cell line lysates probed with mouse mAb to GAPDH (theoretical molecular weight: 36 kDa), NB300-221, dilution 1:2,000: [1] protein standard, [2] HEK293, [3] HeLa, [4] SH-SY5Y, [5] COS1, [6] NIH-3T3, and [7] C6 cells. The GAPDH antibody reveals a single band at ~37 kDa in all cell lines. GAPDH is a house keeping protein, the level of which is relatively unaffected by most experimental manipulations, and, as a result, this antibody has been widely used as a western blotting loading control.

Human eye cell lysate. Antibody used at 1:500. WB image submitted by a verified customer review.

Analysis of GAPDH in chicken pectoralis major using anti-GAPDH antibody. The primary antibody was used at a dilution of 1:4000, incubated for 1 hour at room temperature. Western blot image submitted by a verified customer review.

Application of the approach to other cell lines. The remainder of the protocol from Figure 2 was then applied using 100 ug/mL digitonin as the optimal cytosolic extract in each case. An aliquot of each extract was then analyzed by 4-12% SDS PAGE followed by Western blotting and probing with antibodies to GAPDH, BiP and Histone H3 (Used as an alternative nuclear marker due to weakness of the Lamin A signal in HeLa cells). Image collected and cropped by CiteAb from the following publication (//bmcresnotes.biomedcentral.com/articles/10.1186/1756-0500-2-243) licensed under a CC-BY license.

Endothelial responses provoked by Hb-lipid interactions are attenuated by sulfide donors. Confluence HAoECs were exposed to the obtained Hb samples for 8 hours and VCAM-1 expression was determined by Western blotting. GAPDH were used as a loading control. Representative experiment, n = 3. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/omcl/2018/3812568/), licensed under a CC-BY license.

Regulation of cell migration-associated proteins by NCKU-21 in A549 and CL1-5 cells. Protein expressions of matrix metalloproteinase-2 (MMP-2) and MMP-9 were analyzed in A549 cell treated with NCKU-21 for 24 hr. * P 1.2 to ≤1.3) & high (>1.3) of 3 × MYC-GFP-Bub1 signal & localization in live cells synchronized in mitosis by a thymidine release. Data represent the mean±s.e. of 3 independent experiments, with 58–61 cells measured per condition. Significance was measured for the high group by one-way analysis of variance (ANOVA) & pairwise t-test (Holm–Sidak). (e) Scatter plot of the cytoplasm versus kinetochore GFP levels of individual cells from each of the stable cell lines. The number of cells, R2 (measure of the goodness-of-fit) & significance (one-way ANOVA) are indicated. (f) Western blottings showing levels of the 3 × MYC-GFP-Bub1 proteins in the stable cell lines in whole cell extracts (left) & in cytoplasmic extracts (right). Scale bar, 10 μM. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms9364), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: GAPDH Antibody (1D4) [NB300-221] - Notch1, ApoER2 & Dab1 colocalize in hippocampal neurons. (A) Representative image of fluorescent immunolabeling shows that Notch1 is localized in a subset of CA3 neurons, co-expressing Dab1 & ApoER2 (magnified insert). ApoER2 is highly expressed in all CA3 neurons. (B) Fluorescent immunolabeling shows the distribution of Notch1 & Dab1 as compared to Arc/Arg3.1 in CA3 neurons. (B′) Close up of a dendrite showing colocalization of the three proteins (white arrowheads). (C) Western blot analysis on immunoprecipitated samples from whole hippocampal lysate shows that Notch1 displays a strong interaction with Dab1 whereas Notch1 & ApoER2 interaction appears weaker in the preparation (n = 4 independent experiments). (D) Immunolabeling on immunoprecipitated samples from whole cortical lysates show that Notch1 & Dab1 interact. ApoER2 is bound to both Notch1 & Dab1 (n = 3 independent experiments). (E) Co-IP from synaptosomal membrane fractions reveals a stronger interaction between Notch1 & ApoER2, which is reduced in the N1cKO (n = 3 independent pulldowns). In (C–E) GAPDH is used as a loading control for the inputs & to detect contamination in the IP samples. Scale bar in (A) is 50 μm & (B) 25 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26635527), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: GAPDH Antibody (1D4) [NB300-221] - Reduced expression of UNC5A in FancC−/− cells. a Reduced expression of Unc5A in brain cortex of FancC−/− mice compared to wild-type littermates. Western blotting was performed with the indicated antibodies. Each lane represents a different animal (WT: n = 6; FancC−/−: n = 8). b Bar graph represents ratio of Unc5A normalized to tubulin from WT (n = 6) & FancC−/− mice (n = 8). c Bar graph representation of UNC5A expression normalized to Sdha & Tbp from wild-type (WT) & FancC−/−-derived fibroblasts (n = 3). RLU: Relative light units. d–f HEK293T cells were transfected with UNC5AICD, FANCE & FANCC constructs expressing full-length FANCC (in d), FANCC1–306 (in e) or FANCC307–558 (in f). Constructs were transfected at a molecular ratio of 1:1:1 or with 10 times the molar amount of FANCE, FANCC or UNC5AICD as indicated in the figure. The total amount of transfected plasmid was equalized for all strategies with control empty vectors. Representative immunoblots performed with anti-HA (UNC5AICD), anti-FANCC, anti-FANCE or anti-GAPDH antibodies are shown. Arrows indicate appropriate protein bands. Bar graphs represent the mean fold change ± SEM of UNC5A protein levels normalized to GAPDH compared to 1:1:1 transfection controls in at least 4 separate experiments. *p < 0.05; **p < 0.005 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30213274), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GAPDH Antibody (1D4) [NB300-221] - Depletion of RAD21 & other cohesin subunits leads to differentiation of ESCs.(A): qPCR analysis of esiRNA knock-down efficiency for indicated genes (48 h RNAi, n = 3, error bars denote s.d. & *, ** indicate p<0.05 & 0.01 respectively). Transfection of a non-targeting esiRNA (Luc) was used as a control. Numbers adjacent to the gene names indicate independent, non-overlapping esiRNAs transfected. (B): Western Blot analysis of esiRNA knock-down efficiency for indicated genes (48 h RNAi; esi-1 & esi-2 indicate independent, non-overlapping esiRNAs transfected). Transfection of a non-targeting esiRNA (Luc) was used as a control. GAPDH served as a protein loading control. (C): Alkaline phosphatase staining of ESCs, which had been transfected with esiRNAs targeting RAD21, SMC1a & SMC3 (72 h post RNAi) showed a strong differentiation phenotype compared to a control (Luc). Nanog depletion served as a positive control for ESC differentiation. Scale bars correspond to 100 µm. (D): Quantification of alkaline phosphatase staining (n = 3) separated into undifferentiated (green), mixed (yellow) & differentiated colonies (red). (E+F): qPCR validation of expression changes of (E) stem cell maintenance genes & (F) lineage marker genes upon knock-down of RAD21 (two independent esiRNAs) & Luc control (48 h RNAi, n = 3, error bars denote s.d. & *, **, *** indicate p<0.05, 0.01 & 0.001 respectively). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21589869), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GAPDH Antibody (1D4) [NB300-221] - Application of the approach to other cell lines. (A) HT1080 cells (2 × 105) & HeLa cells (1 × 105) were plated in the wells of a 12 well plate & the digitonin concentration required for optimal extraction of the cytosol was determined as in Figure1. (B) The remainder of the protocol from Figure 2 was then applied using 100 μg/ml digitonin as the optimal cytosolic extract in each case. An aliquot of each extract was then analyzed by 4-12% SDS PAGE followed by Western blotting & probing with antibodies to GAPDH, BiP & Histone H3 (Used as an alternative nuclear marker due to weakness of the Lamin A signal in HeLa cells). (C) Samples were analyzed by 4-12% SDS PAGE followed by staining with Coomassie blue & then silver staining showing the difference in protein composition of each sample. The bracket alongside the gels marks the position of Histones in the RIPA nuclear extract & the asterisk next to the silver stained gels denotes protein bands unique to the E-RIPA extracts. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20003239), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: GAPDH Antibody (1D4) [NB300-221] - Role of the TRIM11-USP14 axis in cell growth & apoptosis. a Levels of endogenous TRIM11 in HCT116 cells grown at 43 °C for 0 or 90 min, or at 43 °C for 90 min, & then at 37 °C for 60 or 120 min. b, c Levels of endogenous TRIM11 (b), or Flag-TRIM11, the indicated proteasome subunits, & USP14 (c) in the SN & P fractions of HCT116 cells with & without heat shock at 43 °C for 90 min. d, g TRIM11 overexpression (d) or knockdown (g) HCT116 cells & the corresponding control cells grown under unstressed, heat shock, & heat shock plus recovery conditions were analyzed for K48 polyUb conjugates & caspase-3 activation. e, f, h Apoptosis in TRIM11 overexpression (e, f) or knockdown (h) HCT116 cells, & the corresponding control HCT116 cells, treated with & without heat shock. i, j Caspase-3 activation (i) & apoptosis (j) in HCT116 cells expressing the indicated proteins and grown under normal or heat stress conditions. k–m K48 polyUb conjugates, caspase-3 activation (k), amyloid-like fibrils (l), & apoptosis (m) in HCT116 cells expressing the indicated proteins and grown under normal & heat stress conditions. In e, f, h, j, l, & m, data represent mean ± SEM (n = 3 unless otherwise indicated). *P < 0.05; **P < 0.01; ***P < 0.001. Uncropped blots are presented in Supplementary Fig. 14 Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-03499-z), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt, Po, Av, Bv, Ca, Ch, ChHa, Dr, Eq, Fe, Ha, Ma-Op, Pm, Rb, Sh, Ze

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Reactivity: Hu, Mu, Rt, Po, Av, Bv, Ca, Ch, ChHa, Dr, Eq, Fe, Ha, Ma-Op, Pm, Rb, Sh, Ze

Applications:

, , , , , , ,

Applications: , , , , , , ,

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Mouse Monoclonal

Host: Mouse Monoclonal

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BSA Free	NB300-221SS

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NB400-105

ABCA1 Antibody - BSA Free

(7 Reviews)

(419 Publications)

0.1 ml

NB400-105

Formalin-fixed paraffin-embedded tissue sections of human prostate cancer were probed for ABCA1 mRNA (ACD RNAScope Probe, catalog # 432291; Fast Red chromogen, ACD catalog # 322360). Adjacent tissue section was processed for immunohistochemistry using rabbit polyclonal (Novus Biologicals catalog # NB400-105) at 1.5ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.

<b>Genetic Strategies Validation and Orthogonal Strategies Validation.</b>HSKMCs were transfected with scrambled siRNA (scr siRNA) or ABCA1 siRNA. The cells were lysed and ABCA1 (A) protein levels were quantified by immunoblotting. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-38014-3) licensed under under a CC-BY license.

<b>Biological Strategies Validation. </b>Analysis of ABCA1 in total cell lysates of RAW264.9 cells treated with vehicle (-) or 9-cisretinoic acid and 22Rhydroxycholesterol (+). Samples used for this testing were 40 ug of total cell post-nuclear lysate from each group.

<b>Biological Strategies Validation. </b>HepG2 cells were grown to 60% confluency, serum starved for 24 hours, and then treated with 1uM TO9 for 24 hours prior to being fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-ABCA1 at 5.0ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Untreated HepG2 cells were grown to 60% confluency, and serum starved for 24 hours prior to being fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-ABCA1 at 5.0ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

ABCA-1 FITC conjugated antibody of human adipose tissue macrophage subsets by flow cytometry. Image from verified customer review.

Association between expression of ABCA1 and survival in ovarian cancer patients. Expression of ABCA1 in 55 ovarian cancer patient samples was determined by IHC in tissue microarray. Representative image of ovarian cancer showing high (left panel) and low (right panel) ABCA1 expression on the cell membrane or cytoplasm (x400). Image collected and cropped by CiteAb from the following publication (//www.clinicalepigeneticsjournal.com/content/7/1/1), licensed under a CC-BY license.

ABCA1 in human primary fibroblasts in culture. 10 ug of total protein. 7.5% TGX gel. Blocking 5% milk in PBST 1h RT. 1:1000 primary ab in BSA 3% PBST O/N at 4C. Secondary 1:5000 HRP 1h RT. Arrow shows around 250 kDa. WB image submitted by a verified customer review.

ABCA1 antibody was tested at 1: 400 in HeLa cells using an Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.

<b>Biological Strategies Validation. </b>Detection of ABCA1 in mouse peritoneal macrophages using NB 400-105 (Lot L). ECL exposure, 1 min. Lane 4: T09 uninduced lysate Lane 5: T09 induced lysate.

Prostates of LXR mutant mice accumulate cholesterol esters through inappropriate LXR target genes regulation. Total protein lysates of WT and LXR null mice under normal or high cholesterol diet were analyzed by western blotting with antibodies against ABCA1, LDLR and ACTIN as a loading control (left panel), quantification of ABCA1 and LDLR protein accumulation levels (right panel). * p<0.05, *** p<0.001 in Student's t test. Error bars represent the +/- mean SEM. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pgen.1003483), licensed under a CC-BY license.

miR-33b reduces cellular cholesterol efflux and serum HDL-C levels. Western blotting for ABCA1 (using NB400-105) and ABCG1 proteins in peritoneal macrophages from WT and KI+/+ mice. Representative images are shown. beta-actin was used as the loading control. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep05312), licensed under a CC-BY license.

miR-33b regulates ABCA1 and SREBP-1. Analysis for ABCA1, SREBP-1, CPT1a, and AMPK alpha protein levels in the livers of WT, KI+/-, and KI+/+ mice. Representative images are shown. TF2B and beta-actin were used as loading controls. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep05312), licensed under a CC-BY license.

Detection of ABCA1 in human prostate epithelium showing luminal and membrane staining.

Liver mRNA & protein expression analysis of chow-fed Osbpl8KO mice.A: qPCR analysis of the quantity of the mRNAs identified at the bottom in chow-fed KO females (open bars) & males (closed bars). The mRNAs were quantified using ribosomal protein 36B4 message as a housekeeping reference. The data are expressed relative to quantity in littermate WT animals of the same gender, & represent mean ± s.e.m. (n = 6; *p<0.05, **p<0.01, T-test). B: Western blot analysis of ABCA1 & SR-B1 proteins in WT & KO mouse liver. The blots were probed with anti-beta -actin as a loading control. Densitometric quantification of the Western blot data is shown on the right. The results were normalized against beta -actin. The data represents mean ± s.e.m. (n = 4). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0058856), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Protein expressions of ABCA1, SR-BI & ABCG1 in liver & macrophage by Western blot. Simvastatin increased the expressions of ABCA1 & ABCG1 in liver & ABCA1 in macrophage, Both L-4F & the combination group improved the expressions of ABCA1, SR-BI & ABCG1 in liver & ABCA1 & ABCG1 in macrophage. 1P < 0.05, 2P < 0.001, vs. AS group; aP < 0.05, bP < 0.001, vs. Simva group;*P < 0.05, **P < 0.001, vs. L-4F group. Image collected & cropped by CiteAb from the following publication (//lipidworld.biomedcentral.com/articles/10.1186/1476-511X-12-180), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

ABCA1, ABCG1 & SR-BI protein expressions. a & b Hepatic protein expressions of ABCA1, ABCG1 & SR-BI were significantly decreased in COMT−/− mice at GD 18.5, compared to C57BL/6 J mice. Decreased hepatic ABCA1 expression was also observed at 10 days postpartum. ATI-5261 increased ABCA1 & ABCG1 expression in the liver at 10 days postpartum. c Placental protein expressions of ABCA1 & ABCG1 was reduced in COMT−/− mice, compared to C57BL/6 J mice. ATI-5261 treatment significantly increased ABCA1 levels in the placenta of COMT−/− mice. d Representative immunoblots of the corresponding proteins in the placenta with mouse RAW264.7 cell lysate included as positive control. Similar results were obtained when the experiment was repeated with lysates prepared from three batches of tissues. Data are presented as mean ± SEM. Groups (n = 8 in all groups) were compared using one-way ANOVA with post-hoc analysis (Tukey’s procedure). *, p < 0.05 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30237900), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Effects of 13-HODE & LA in the presence & absence of PPAR alpha & PPAR gamma selective antagonists on molecular markers of cholesterol homeostasis in RAW264.7 macrophages. RAW264.7 cells were pre-treated without or with the PPAR alpha selective antagonist GW6471 or the PPAR gamma selective antagonist GW9662 & subsequently treated without (vehicle control) or with 2.5 μmol/L 13-HODE or 100 μmol/L LA for 24 h. Afterwards, cells were lysed & subsequently processed for western blotting as described in the materials & methods section. A, Representative immunoblots specific for ABCA1, ABCG1, SR-BI, LXR alpha , & beta -actin which was used for normalization are shown. B, Bars represent data from densitometric analysis & are means ± SD from three independent experiments (n = 3). Data are expressed as percentage of protein concentration of vehicle control cells. Results from statistical analysis are indicated: Significant effects are denoted with superscript letters. Bars marked without a common superscript letter significantly differ (P < 0.05). Image collected & cropped by CiteAb from the following publication (//lipidworld.biomedcentral.com/articles/10.1186/1476-511X-10-222), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - (A) Upregulation of ABCA1 & ABCG1 & (B) enhancement of cholesterol efflux by purple Perilla frutescens extracts (PPE) in 50 μg/ml Cu2+-oxidized low-density lipoproteins (LDL)-exposed J774A.1 murine macrophages. (A) For the measurement of ABCA1 & ABCG1 expression, total cell lysates were subjected to western blot analysis with a primary antibody against ABCA1 or ABCG1. beta -actin was used as an internal control. Bar graphs (means ± SEM, n=3) represent quantitative densitometric results of the upper bands. (B) Cholesterol efflux was expressed as the percentage of fluorescence in the medium relative to the total fluorescence. Bar graphs denoted without a common letter indicate significant difference, P<0.05. Image collected & cropped by CiteAb from the following publication (//www.spandidos-publications.com/10.3892/ijmm.2015.2101), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - 6‐Dihydroparadol increases A) ABCA1 & B) ABCG1, but not C) SR‐BI protein levels in cholesterol‐loaded THP‐1‐derived macrophages. THP‐1 cells were differentiated as described in Figure 2, & then loaded with unlabeled cholesterol for 24 h. Cells were treated with increasing concentrations of 6‐dihydroparadol (1–30 μm) for another 24 h. The protein levels of ABCA1, ABCG1, & SR‐B1 were detected by Western blot analysis. The control was treated with solvent vehicle (0.1% DMSO). As a positive control, TO901317 (5 μm, 24 h) was used. The bar graphs represent mean ± SD from three independent experiments. *p < 0.05, **p < 0.01, & ***p < 0.001 versus control (determined by Student's t‐test or ANOVA with Bonferroni's post hoc test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29802792), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Akt inhibition suppresses mTORC1 activity.BHK-ABCA1 cells were induced overnight with mifepristone (10 nM) & then incubated with DEBC, LY294002 or rapamycin at indicated concentrations for 2 h. Cells were then lysed & analyzed for ABCA1 & phosphorylated S6K by Western blotting. Actin was also blotted as loading control. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0113789), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Reversal of hepatic steatosis by reduction of SREBP-1 levels.(a) Western blotting analysis of SREBP-1 & ABCA1 levels in livers of miR-33+/+Srebf1+/+, miR-33+/+Srebf1+/−, miR-33−/−Srebf1+/+ & miR-33−/−Srebf1+/− mice. Representative western blot images are shown (n=4). Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/24300912), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - 6‐Dihydroparadol increases A) ABCA1 & B) ABCG1 protein stability. THP‐1 cells were differentiated as described in Figure 2. Then cells were loaded with unlabeled cholesterol & treated with 6‐dihydroparadol (10 μm), or solvent vehicle (0.1% DMSO, control) for 24 h. Cells were lysed at different time points (0, 1, 2, 3, 4, & 6 h) after treatment with the protein synthesis inhibitor cycloheximide (CHX, 100 μm). The protein levels of both ABCA1 & ABCG1 were detected by Western blot analysis. The data points represent mean ± SD from three independent experiments. *p < 0.05 & **p < 0.01 versus control at the same time point (determined by Student's t‐test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29802792), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Ligand activation of LXR inhibits LPS-induced MAP kinase activation through Abca1 induction.(A–D) Bone marrow-derived macrophages from Lxr alpha −/−Lxr beta −/− & control wild-type mice (A, B), or bone marrow-derived macrophages from myeloid-specific Abca1−/− & control wild-type mice (C, D) were pretreated with GW3965 (1 µM) overnight, followed by stimulation with LPS (10 ng/ml) for 20 min or 1 hr. Whole cell lysates were harvested & protein expression was analyzed by immunoblotting with the indicated antibodies (A, C). Protein expression was quantified by Image Quant TL7.0 (B, D). N = 4–6 per group. *p < 0.05, **p < 0.01, NS, not significant. Error bars represent means ± SEM.DOI://dx.doi.org/10.7554/eLife.08009.012 Image collected & cropped by CiteAb from the following publication (//elifesciences.org/articles/08009), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Western Blotting for ABCA1, as detailed in Supplementary Protocol #5 (Additional file 1). Caco-2 cells cultivated on filter inserts were treated for 48 h with bexarotene (RXR agonist, 5 μM), T0901317 (LXR agonist, 10 μM), GW3965 (LXR agonist, 10 μM) or the vehicle control (DMSO 0.1%). Protein expression was then determined by western blot analysis. Results were normalized for interexperimental variance & then set in relation to the vehicle control. Bar graphs represent mean ± SD from three independent experiments. A representative blot is shown - the bands stem from the same membrane & the vertical line indicates excised irrelevant bands. ns not significant, *P < 0.05, **P < 0.01 versus solvent vehicle (determined by paired t-test) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32308567), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Expression of ABCA1 & CROT in livers & RIP140 in macrophages is elevated in miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice. A, Quantitative real-time PCR analysis of Abca1, Crot, Cpt1a, & Prkaa1in livers from miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. Values from miR-33+/+Apoe−/− mice were set at 100%. Values are mean±SE (n=9 to 11each); **P<0.01. B, Western analysis of ABCA1, CROT, CPT1a, & AMPK alpha in livers from miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. beta -actin was used as a loading control. C, Densitometry of ABCA1, CROT, CPT1a, & AMPK alpha in livers from miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. Values from miR-33+/+Apoe−/− mice were set at 100%. Values are mean±SE (n=4 each); *P<0.05. D, Total cholesterol, free cholesterol, cholesterol ester, & triglyceride levels in livers of miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. Values are mean±SE (n=9 to 11 each). E, HE staining of livers of miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice at age 20 weeks fed NC. Scale bar: 100 μm. F, Quantitative real-time PCR analysis of Nrip1 (RIP140) in peritoneal macrophages from miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. Values from miR-33+/+Apoe−/− mice were set at 100%. Values are mean±SE (n=7 each). G, Western analysis of NRIP1 (RIP140) in peritoneal macrophages from miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. GAPDH was used as a loading control. H, Densitometry of NRIP1 (RIP140) in peritoneal macrophages from miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. Values from miR-33+/+Apoe−/− mice were set at 100%. Values are mean±SE (n=4 each); *P<0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23316322), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Basal ABCA1 expression & function in patient-derived iRPEs. (A) Representation of patient-derived iPSC lines & genotypes harboring polymorphisms in ABCA1 that are associated with decreased or increased risk for AMD development. (B) Relative expression of ABCA1 mRNA in patient-derived iRPEs under basal conditions normalized to RPL28 & decreased risk group. (C) Western blot analysis of ABCA1 levels in patient-derived iRPEs under basal conditions. Actin was detected as loading control. ABCA1 expression was quantified & normalized to actin. (D,E) Relative expression of ABCG1 (D) & NR1H3 (E) mRNA in patient-derived iRPEs under basal conditions normalized to RPL28 & decreased risk group. (F) Cholesterol efflux in patient-derived iRPEs after direct cell labeling & in the presence of ApoAI. (G) Cholesterol efflux in patient-derived iRPEs after phagocytosis of BODIPY-cholesterol-loaded POSs & in the presence of ApoAI. (H) Quantification of phagocytosed POSs per nuclei in patient-derived iRPEs. Values shown are means ± SD (n = 3). * p < 0.05; *** p < 0.001. Unpaired Student’s t-test. (I) Representative fluorescence microscopy images of phagocytosed FITC-labeled POSs (green) & staining for ZO-1 (red) & DAPI (blue) of increased risk cell line IPS19-00096. Scale bar = 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35328615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - A–C) Effects of 6‐dihydroparadol & specific proteasomal, lysosomal, & calpain inhibitors on the ABCA1 protein levels. THP‐1 cells were differentiated as described in Figure 2, & then loaded with unlabeled cholesterol for 24 h. Cells were treated with or without 6‐dihydroparadol (6‐DP, 30 μm) for 24 h & incubated for another 3 h with or without the proteasome inhibitor lactacystin (10 μm), the lysosomal inhibitor chloroquine (100 μm), or the calpain inhibitor calpeptin (30 μg mL−1). The ABCA1 protein levels were detected by Western blot analysis. The bar graphs represent mean ± SD from three independent experiments. *p < 0.05, **p < 0.01, & ***p < 0.001 versus control; n.s., not significant versus control (determined by Student's t‐test or ANOVA with Bonferroni's post hoc test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29802792), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Suppressive effects of Ucn1 on foam cell formation & related protein expression in human monocyte-derived macrophages.Human monocytes were incubated for 7 days with RPMI-1640 supplemented with 10% human serum & the indicated concentrations of Ucn1, followed by a 19 h-incubation with 50 µg/ml oxLDL in the presence of 0.1 mmol/l [3H]oleate. Intracellular CE accumulation was determined from the radioactivity of cholesterol-[3H]oleate. Otherwise, before the addition of oxLDL, cells were harvested & subjected to immunoblotting analyses for CD36, ACAT1, or ABCA1. beta -Actin served as a loading control. Data are expressed as means ± SEM from 4–6 independent experiments with monocytes from 4–6 different donors. Baseline (1 fold) = 8.4±2.2 nmol/mg cell protein. *P<0.05, †P<0.001 vs. 0 nmol/l of Ucn1. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0110866), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Effect of evodiamine on ABCA1 transcription & the degradation rate of ABCA1 protein. (a) Differentiated THP-1 macrophages were incubated with 10 μM evodiamine or 10 μM pioglitazone as positive control for 24 h. Total RNA was extracted & ABCA1 mRNA expression levels were quantified by qRT-PCR. (b) 293 T cells were transfected with a luciferase reporter construct driven by the human ABCA1 promoter as described in the Materials & Methods section. After transfection, cells were treated with 10 μM Evodiamine or 1 μM T0901317 as positive control for 24 h. (c) Differentiated THP-1 macrophages were incubated for 24 h with (black circle) or without (Veh; white circle) evodiamine (10 μM) & lysed after addition of cycloheximide (CHX; 140 μM) at different time points (0, 10, 20, 40 min). Western blot analysis shows the decline of ABCA1 protein level with cycloheximide in the presence & absence of evodiamine. All data are means ± S.D. (n = 3) vs. solvent vehicle control (DMSO), *p < 0.05, **p < 0.01, n.s. no significance (ANOVA/Bonferroni). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30038271), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - miR-33 deficiency improved cholesterol efflux in macrophages. A, Quantitative real-time PCR analysis of Abca1 & Abcg1 in macrophages from miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. Values from miR-33+/+Apoe−/− were set at 100%. Values are mean±SE (n=7 each); ***P<0.001. B, Western blotting analysis of ABCA1 & ABCG1 in thioglycollate-elicited peritoneal macrophages from miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. GAPDH was used as a loading control. C, Cholesterol efflux from thioglycollate-elicited peritoneal macrophages in the presence or absence of apoA-I (5 or 10 μg/mL). Values are mean±SE (n=6 each); **P<0.01. D, Cholesterol efflux from thioglycollate-elicited peritoneal macrophages in the presence or absence of HDL-C (50 or 100 μg/mL). Values are mean±SE (n=6 each); **P<0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23316322), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Srebf1 is a miR-33 target gene.(a) Relative changes in lipid metabolism-related genes in the livers of miR-33−/− mice compared with miR-33+/+ mice fed NC at 16 weeks of age. (n=5–8 each,*P<0.05 in Student’s t-test). (b) Conservation of miR-33 target regions in the 3′UTR of Srebf1. Underlined sequences are the potential binding site of miR-33 seed sequences. * indicates the conservation among spieces. (c) 3′UTR reporter assay used to verify the target. Luciferase reporter activity of human & mouse SREBP-1 gene 3′UTR constructs in 293T cells overexpressing miR-control (miR-Con) & miR-33 (n=4 each, *P<0.05 & ***P<0.001 in Student’s t-test). (d) miR-33 dose-dependent changes in luciferase reporter activity of mouse Srebf1 3′UTR construct in 293T cells. miR-Con & miR-146a is used as a negative control (n=4 each, *P<0.05 & ***P<0.001 in one-way analysis of valiance test). (e) Luciferase reporter activity of the WT or mutant Srebf1 3′UTR at the potential miR-33 binding site in 293T cells (n=4 each, **P<0.01 in Student’s t-test). (f,g) Luciferase reporter activity of SRE-promoter (f) or FAS-promoter (g) in 293T cells. 293T cells were co-transfected with mouse Srebf1 with the full-length 3′UTR or without the 3′UTR, along with expression plasmids for miR-negative control, or miR-33. Values are the mean±s.e. (n=4 each, **P<0.01 versus miR-Con. ***P<0.001 versus miR-Con in one-way analysis of valiance test). (h) Western blotting analysis of SREBP-1, ABCA1, & IRS-2 in miR-33 transduced HepG2 cells & primary hepatocytes & hepatocytes prepared from miR-33+/+ & miR-33−/− mice. Representative western blot images are shown (n=4). Values are the means±s.e.m. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24300912), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - (A) Chemical structure, (B) cytotoxicity of alpha -asarone, (C) enhancement of cholesterol efflux by alpha -asarone, (D & E) upregulation of ABCA1 & ABCG1 by alpha -asarone & beta -asarone, & (F) elevation of retinoid X receptor (RXR) alpha transcription. J774A.1 murine macrophages were exposed to 50 μg/ml oxidized low-density lipoprotein (LDL) & treated with 1–10 μM purple Perilla frutescens extracts (PPE)-alpha -asarone & 5–10 μM beta -asarone. (B) MTT assay was performed for the measurement of alpha -asarone toxicity. Graph data represent 1 of 4 independent experiments with multiple estimations. Values are expressed as the percentage cell survival relative to the untreated control cells (cell viability, 100%). (C) Cholesterol efflux was expressed as the percentage fluorescence in the medium relative to total fluorescence. (D & E) For the measurement of ABCA1 & ABCG1 expression, total cell lysates were subjected to western blot analysis with a primary antibody against ABCG1 or ABCG1. beta -actin was used as an internal control. Bar graphs (means ± SEM, n=3) represent quantitative densitometric results of the upper bands. Bar graphs denoted without a common letter indicate significant difference, P<0.05. (F) RXR alpha mRNA expression was measured by RT-PCR. GAPDH was used as a housekeeping gene for the co-amplification with RXR alpha . Image collected & cropped by CiteAb from the following publication (//www.spandidos-publications.com/10.3892/ijmm.2015.2101), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Bilirubin significantly decreases expression of ATP‐binding cassette transporter A1 (ABCA1) protein in THP‐1 macrophages treated with LXR agonist (TO901317) to upregulate ABCA1 protein. THP‐1 cells were differentiated for 72 hours with 200 nmol/L phorbol‐12‐myristate‐13‐acetate & then loaded with unlabeled cholesterol for another 24 hours. Cells were treated with TO901317 (5 μmol/L) or solvent vehicle (0.1% dimethyl sulfoxide [DMSO]) for 24 hours. Afterward, cells were treated with bilirubin (17.1 μmol/L) or solvent vehicle (0.1% DMSO) for 16 hours. The protein levels of ABCA1 were detected by western blot analysis. The bar graphs present mean±SD from 3 independent experiments. ***P<0.001 vs control (determined by ANOVA with Bonferroni post hoc test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28455345), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Induction of cholesterol efflux attenuates the enhancement in HIV-1infectivity in NPCD55 cells. (A) Normal & NPCD55 cells were treated with 5 μM TO-901317 at 24 h post-infection & cultured for 72 h in the presence of the compound. At 96 h post-infection the cells were harvested & ABCA1, Gag, & beta -actin expression was detected by Western blotting analysis. All samples were loaded on the same gel. (B) AmplexRed assay was performed to measure free cholesterol content of untreated (filled) & TO-901317-treated (slashed) cells. Cholesterol content was normalized to protein concentration. (C) AmplexRed assay was performed to measure virion-associated cholesterol from purified virus produced in untreated (filled) & TO-901317-treated (slashed) infected cells. Virion-associated cholesterol content was normalized to p24 concentration. (D) TZM-bl reporter assay was performed to measure virus infectivity when cholesterol efflux was induced via TO-901317 stimulation. The ρ values were calculated by performing student T-test (*denotes < 0.05). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22273177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Effects of apabetalone on ABCA1 expression & carboplatin response in ovarian cancer cells. (A) Apabetalone treatment reduced expression of ABCA1 in OVCAR-5 & OVCAR-5 CBPR cell lines. Cells were treated for 72 h with control medium (DMSO) or apabetalone (80 µM). Protein extracts from OVCAR-5 (~30 µg) were electrophoresed & immunoblotted with rabbit polyclonal ABCA1 antibody (1/1000, NB400-105, Novus Biological) & beta -actin (1/2000, Abcam) was used as a loading control. (B) Quantification of Western blot. OVCAR-5 (C), CaOV3 (D), OVCAR-5 CBPR (E) & CaOV3 CBPR (F) cell survival following treatment with carboplatin (CBP) alone (0-200 µM, black line) & in combination with apabetalone (Apa, 80 µM, purple line). (G) Carboplatin IC50 for OVCAR-5 & OVCAR-5 CBPR cells ± apabetalone. Data are mean ± SEM from three independent experiments. *P < 0.05, Student’s t-test. (H) Carboplatin IC50 for CaOV3 & CaOV3 CBPR cells ± apabetalone. Data are mean ± SEM from three independent experiments. P < 0.05, Student’s t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35582032), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Expression of ATP‐binding cassette transporter A1 (ABCA1) protein in THP‐1 macrophages treated with bilirubin & in peripheral blood mononuclear cells (PBMCs) from Gilbert syndrome (GS) patients. A, Bilirubin suppresses the expression of ABCA1 protein in THP‐1‐derived macrophages time‐dependently. THP‐1 cells were differentiated as described in Figure 1 & then loaded with unlabeled cholesterol for another 24 hours. Cells were treated with bilirubin (17.1 μmol/L) for 4, 8, 16, & 24 hours. The protein levels of ABCA1 were detected by western blotting. Control was treated with solvent vehicle (0.1% dimethyl sulfoxide). The bar graph presents mean±SD from 3 independent experiments. B, Expression of ABCA1 protein is decreased in PBMCs from participants with high bilirubin blood levels (GS) compared with healthy controls. The protein levels of ABCA1 were detected by western blotting. The bar graph presents mean±SEM (n=28 per group). *P<0.05, **P<0.01 & ***P<0.001 vs control. ns indicates not significant (determined by Student t test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28455345), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - (A) Time course response of ABCA1 induction by pioglitazone, & (B) enhancement of ABCA1 & (C) peroxisome proliferator-activated receptor gamma (PPAR gamma ) by purple Perilla frutescens extracts (PPE). J774A.1 murine macrophages were incubated with 10 μM pioglitazone & 50 μg/ml oxidized low-density lipoprotein (LDL) in the absence or presence of 1–10 μg/ml PPE. For the measurement of expression of (A & B) ABCA1 & (C) PPAR gamma , total cell lysates were subjected to western blot analysis with a primary antibody against ABCA1 or PPAR gamma . beta -actin was used as an internal control. Bar graphs (means ± SEM, n=3) represent quantitative densitometric results of the upper bands. Bar graphs denoted without a common letter indicate significant difference, P<0.05. Image collected & cropped by CiteAb from the following publication (//www.spandidos-publications.com/10.3892/ijmm.2015.2101), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - LXR agonist-stimulated ABCA1 expression & function in patient-derived iRPEs. Patient-derived iRPE lines were stimulated for 16 h with LXR agonist & are relative to unstimulated cells shown in Figure 2. (A) Expression of ABCA1 mRNA normalized to RPL28 & relative to unstimulated cells shown in Figure 2B. (B) Western blot analysis of ABCA1 protein levels. Actin was detected as loading control. ABCA1 expression was quantified & normalized to actin. (C) Cholesterol efflux assay after direct cell labeling & in the presence of ApoAI. (D) Cholesterol efflux assay after phagocytosis of BODIPY-cholesterol-loaded POSs & in the presence of ApoAI. Data are relative to unstimulated cells shown in Figure 2F,G, respectively. Data are presented as means ± SD (n = 3). Unpaired Student’s t-test. * p < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35328615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Akt inhibition by LY294002 or Akt1/2 also enhances cholesterol efflux to apoA-I from ABCA1-expressing BHK cells.A) BHK cells were induced as in Fig. 1 & then incubated with LY294002 (200 µM) for 2 h. Cells were then lysed & analysed for ABCA1, phosphorylated Akt (p-Akt) & total Akt by Western-blotting. Hsp70 was also blotted as loading control. B & C) BHK cells were labeled with [3H] cholesterol & induced with 10 nM mifepristone overnight. Cholesterol efflux was measured after 2 h incubation with BSA (1 mg/ml) or BSA plus apoA-I (5 µg/ml). Some of cells were also incubated with indicated doses of LY294002 (B) or Akt1/2 (C), in addition to apoA-I, during 2 h efflux period. Results are presented as the average of triplicate wells with standard deviation & representative of more than three independent experiments. *** P<0.0001, **P<0.001 & *P<0.05 vs apoA-I only. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0113789), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Generation, differentiation & analysis of ABCA1-deficient iRPEs. (A) Immunofluorescence staining for ZO-1 (red) in 4-week cultured iRPEs. Nuclei were stained with DAPI (blue). Scale bar = 100 μm. (B) Bright-field microscopy of iRPEs. (C) Ct values of RPE marker genes BEST1, OTX2, RLBP1 & RPE65 in iRPEs & parental iPSCs obtained by qPCR. Values shown are means ± SD (n = 3). Unpaired Student’s t-test. **** p < 0.0001. (D) Sequence alignment of parts of exon 14 (top) & exon 46 (bottom) of ABCA1-deficient iRPE cell clones & parental line. (E) Western blot of ABCA1 protein levels in ABCA1-deficient iRPE cell lines & parental cell line after 16 h of stimulation with 1 μM LXR agonist. Actin was detected as loading control. (F) Relative expression of ABCA1 mRNA in ABCA1-deficient iRPE cell lines & parental cell line normalized to ACTB. Values shown are means ± SD (n = 3). One-way ANOVA with Tukey’s post hoc test. **** p < 0.0001. (G) Relative expression of ABCG1 mRNA in ABCA1-deficient iRPE cell lines & parental cell line normalized to ACTB. Values shown are means ± SD (n = 3). One-way ANOVA with Tukey’s post hoc test. ** p < 0.01; *** p < 0.001. (H) Cholesterol efflux assay in iRPEs in the presence of ApoAI and/or LXR agonist (LXR ag). Values shown are means ± SD (n = 3). Two-way ANOVA with Bonferroni post hoc test vs. DMSO + BSA control. **** p < 0.0001. (I) Bright-field (BF) microscopy & Nile Red fluorescence microscopy (overview & close-up view marked with white squares) of 4-week cultured ABCA1-deficient iRPE cell lines & parental cell line. Relative fluorescence was quantified & is shown as mean ± SD (n = 8). One-way ANOVA with Tukey’s post hoc test. * p < 0.05, **** p < 0.0001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35328615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Akt inhibition enhances cholesterol efflux to apoA-I from wt MEFs, but not from TSC2-/- MEFs where mTORC1 is constitutively activated.A) Mouse embryonic fibroblasts (MEFs), wt & TSC2-/-, were induced overnight with T0901317 (10 µM). Cell lysates were collected for Western blotting for phosphorylated S6K & total S6K. B) wt & TSC2-/- MEFs were incubated with or without T0901317 (10 µM) overnight. Some of cells were then incubated with apoA-I (10 µg/ml) or apoA-I plus DEBC (25 µM) for 2 h. The expression of ABCA1, phosphorylated Akt, total Akt & loading control hsp70 were analyzed by Western blotting. C) wt & TSC2-/- were labeled with [3H] cholesterol for 1 day & then incubated with or without T0901317 (10 µM) overnight. Some of cells were then incubated with apoA-I (10 µg/ml) or apoA-I plus DEBC (25 µM) for 2 h to analyze cholesterol efflux. Data presented as the average of triplicate wells with standard deviation & representative of at least three independent experiments. *** P<0.0001 vs apoA-I only. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0113789), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Akt inhibition by DEBC enhances cholesterol efflux to apoA-I specifically from ABCA1-expressing BHK cells.A) BHK-ABCA1 cells were induced with mifepristone (10 nM) overnight & then incubated with indicated doses of DEBC for 2 h. Cells were then lysed & Western-blotted for ABCA1, phosphorylated Akt (p-Akt) & total Akt. Hsp70 was also blotted as loading control. B) BHK-ABCA1 cells were labeled with [3H] cholesterol & induced overnight as above. After 2 h incubation with BSA (1 mg/ml) or BSA plus apoA-I (5 µg/ml), cholesterol efflux was measured as described in the Methods section. Some of the cells were also incubated with indicated doses of DEBC, in addition to apoA-I, during 2 h efflux period. C) BHK-ABCA1 & BHK-A937V cells were induced with mifepristone (10 nM) overnight. 2 h Cholesterol efflux was measured as above either with or without DEBC (25 µM). Results are presented as the average of triplicate wells with standard deviation & representative of more than three independent experiments. *** P<0.0001 & **P<0.001 vs apoA-I only. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0113789), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - LXRs control expression of genes involved in cholesterol homeostasis & fatty acid synthesis in MPECs.(A) qPCR analysis of Abca1, Abcg1 & Idol levels in WT (+/+) & Lxr alpha beta −/− (lxr−/−) MPECs after DMSO (vehicle) or T0901317 stimulation (B) Effect of 9-cis retinoic acid and/or T0901317 stimulation on Abca1 accumulation levels in WT (+/+) & Lxr alpha beta −/− (lxr−/−) MPECs (C) qPCR analysis of Srebp1, Acc, & Fas levels in WT (+/+) & Lxr alpha beta −/− (lxr−/−) MPECs after DMSO (vehicle) or T0901317 stimulation. (qPCR analysis results from 4 independant experiments & was normalized using 36b4 gene). *p<0.05, **p<0.01, ***p<0.001 in Student’s t test. Error bars represent mean ± SEM. (D) Western blot analysis was performed on WT (+/+) or Lxr alpha beta −/− MPECs using Srebp1c, Abca1 & beta -Actin antibodies. (E) Oil-Red O staining (ORO) & Normarski/Dapi of WT (+/+) & Lxr alpha beta −/− MPECs, or WT MEFs, treated for 48h with DMSO (vehicle) or T0901317 (1 µM). Head arrows indicate lipid droplets. Scale bars 100 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23554947), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Anti-ApoA-1 IgGs upregulate ABCA1. ABCA1 was increased after 16 & 24 h of anti-ApoA-1 IgG exposure to HMDM at the mRNA level as revealed by RT-PCR (a) as well as at the protein level after 24 h anti-apoA-1 IgG stimulation, as revealed by Western blot analysis (b). In panel a, data are expressed as fold change expression of the mean ± SD of ABCA1 calculated by delta delta CT method of six independent experiments (n = 6) & values were normalized to untreated condition. p-values were calculated using the Student’s t-test: * p < 0.05, ** p < 0.01, **** p < 0.0001. (b) One of four representative Western blots is shown. Data are the mean ± SD of band intensity volume/actin intensity volume of four independent experiments (n = 4). p-values were calculated using the Student’s t-test: * p = 0.02, ** p = 0.005. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31766415), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - ABCA1 protein expression is upregulated in ovarian cancer cells with acquired carboplatin resistance. ABCA1 expression in OVCAR-5 (A) & CaOV3 (C) & carboplatin resistant OVCAR-5 CBPR (B) & CaOV3 CBPR (D) by immunocytochemistry using rabbit polyclonal ABCA1 antibody (1/100, NB400-105, Novus Biological). (E) OVCAR-5 cells with Rabbit IgG & (F) CaOV3 cells with Rabbit IgG. (G) Protein extracts from OVCAR-5 (~30 µg) & CaOV3 cell lines (~60 µg) were electrophoresed & immunoblotted with rabbit polyclonal ABCA1 antibody (1/1000, NB400-105, Novus Biological), & beta -actin (1/2000, Abcam) was used as a loading control. A major band was detected at ~250 kDa, which is the predicted size for ABCA1. (H) Quantitation of ABCA1 Western blots. Data are from 2-4 independent experiments. Statistical significance was determined using the Student’s t-test, *P < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35582032), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Evodiamine enhances apo A1-mediated ChE from THP-1 macrophages & increases ABCA1 protein level. (a) Differentiated THP-1 cells were loaded with [3H]-cholesterol together with the indicated treatments for 24 h. On the next day, the cells were washed twice with PBS & incubated with the same compounds [solvent vehicle control (Veh; ≤0.1% DMSO), evodiamine (1–20 μM), & the PPAR gamma agonist pioglitazone (10 μM) as positive control] with or without 10 µg/mL apo A1. Extracellular as well as intracellular radioactivities were quantified with scintillation counter. Differentiated THP-1-derived macrophages were treated with solvent vehicle control (Veh; ≤0.1% DMSO), evodiamine (10 μM), & the PPAR gamma agonist pioglitazone (10 μM) as positive control. After 24 h incubation, the cells were lysed & 20 μg protein was resolved via SDS-PAGE. Immunodetection was performed with antibodies against the indicated proteins, ABCA1 (b), ABCG1 (c), & SR-B1 (d), & visualized by chemiluminescence detection. All experiments were performed at least three times & data are presented as means ± S.D. vs. solvent vehicle control, *p < 0.05, **p < 0.01, ***p < 0.001, n.s. no significance (ANOVA/Bonferroni). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30038271), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Functional miR-33 responsive elements in the 3′UTR of ATP8B1 & ABCB11A,B. Conserved sequences in the 3′UTR of ATP8B1 & ABCB11 are partially complementary to miR-33. The element in human ATP8B1 is located 1877–1897 nt after the stop codon. The element in ABCB11 overlaps the stop codon in primates, while rodents show a conserved sequence 732–751 nt after the stop codon.C,D. Luciferase assays in HEK293 cells using the whole 3′UTR of human or murine ATP8B1 & ABCB11, or a single copy of the responsive elements (RE) identified above, or mutant responsive elements (RE*; where AATGCA was mutated to GGGTTG to prevent complementarity to the seed sequence of the miRNA), co-transfected with (closed bars) or without (open bars) a vector to overexpress miR-33. In grey, data from empty (negative control) & R33 (positive control containing a 100% match to miR-33) reporter vectors.E,F. Relative mRNA expression of canalicular transporters in primary murine hepatocytes (n = 4 dishes/condition) & human HuH-7 hepatoma cells (n = 3 dishes/condition) transduced 48 h with empty or miR-33 adenovirus.G. Relative protein levels in HuH7 cells transduced with empty or miR-33 adenovirus. Some cells were incubated for 16 h in the presence of FXR:RXR agonists (2 µmol/L GW4064 : 1 µmol/L 9-cis-retinoic acid) to induce ABCB11. Asterisk indicates a non-specific band. Data shown as mean ± SD; **p < 0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22767443), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Gas6 treatment enhances expression of LXR alpha & LXR beta & their target genes in BMDM.Mouse BMDM were stimulated with 400 ng/ml Gas6, 1 μM T0901317, 10 ng/ml interferon (IFN)-gamma , 10 ng/ml IL-4, or 100 ng/ml LPS for 4 h (a,b) or 400 ng/ml Gas6 for the indicated times (c–g). (a,b,e) The amounts of the LXR alpha , LXR beta , ABCA1, ABCG1, ApoE, AIM, Arg2, VEGF, YM1, & Arg1 mRNAs were analyzed by real-time PCR & normalized to that of Hprt mRNA. (c,d,f,g) The relative abundances of LXR alpha , LXR beta , ABCA1, ABCG1, ApoE, Aim, Arg2, & VEGF proteins were determined by Western blotting analysis. The relative densitometric intensity was determined for each band & normalized to beta -actin. Data in all bar graphs are means ± SEM of three independent experiments. *P < 0.05 compared with control. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27406916), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - (A) Time course response of ABCA1 induction by TO-091317 & (B) upregulation of ABCA1 by purple Perilla frutescens extracts (PPE), & (C) enhancement of liver X receptor (LXR) alpha induction by PPE. J774A.1 murine macrophages were cultured with 1 μM TO-091317 or 50 μg/ml Cu2+-oxidized low-density lipoprotein (LDL) in the absence or presence of 1–10 μg/ml PPE. For the measurement of expression of (A & B) ABCA1 & (C) LXR alpha , total cell lysates were subjected to western blot analysis with a primary antibody against ABCA1 or LXR alpha . beta -actin was used as an internal control. Bar graphs (means ± SEM, n=3) represent quantitative densitometric results of the upper bands. Bar graphs denoted without a common letter indicate significant difference, P<0.05. Image collected & cropped by CiteAb from the following publication (//www.spandidos-publications.com/10.3892/ijmm.2015.2101), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ca, Ch, ChHa, Eq, Ha, Md, Pm, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca, Ch, ChHa, Eq, Ha, Md, Pm, Rb

Applications:

, , , , , , , , , , , , , , , ,

Applications: , , , , , , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-105AF350
Alexa Fluor 405	NB400-105AF405
Alexa Fluor 488	NB400-105AF488
Alexa Fluor 532	NB400-105AF532
Alexa Fluor 594	NB400-105AF594
Alexa Fluor 647	NB400-105AF647
Alexa Fluor 700	NB400-105AF700
Alexa Fluor 750	NB400-105AF750
Allophycocyanin	NB400-105APC
Allophycocyanin/Cy7	NB400-105APCCY7
Biotin	NB400-105B
CoraFluor 1	NB400-105CL1
DyLight 350	NB400-105UV
DyLight 405	NB400-105V
DyLight 488	NB400-105G
DyLight 550	NB400-105R
DyLight 594	NB400-105DL594
DyLight 650	NB400-105C
DyLight 680	NB400-105FR
DyLight 755	NB400-105IR
FITC	NB400-105F
HRP	NB400-105H
Janelia Fluor 525	NB400-105JF525
Janelia Fluor 549	NB400-105JF549
Janelia Fluor 646	NB400-105JF646
Janelia Fluor 669	NB400-105JF669
PE	NB400-105PE
PE/Atto594	NB400-105PEATT594
PE/Cy5.5	NB400-105PECY55
PE/Cy7	NB400-105PECY7
PerCP	NB400-105PCP
mFluor Violet 450 SE	NB400-105MFV450
mFluor Violet 500 SE	NB400-105MFV500
mFluor Violet 610 SE	NB400-105MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-105SS

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NB400-104

SR-BI Antibody - BSA Free

(5 Reviews)

(243 Publications)

0.1 ml

NB400-104

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-104] - SR-BI antibody was tested in human fibroblast samples fixed in 4% PFA and permeabilized in PBS (0.2% Tween). Primary incubation overnight at 4C using a 1:100 dilution in PBS (0.1% Tween) with 1% BSA. Secondary antibody is anti-rabbit conjugate to Alexa Fluor 488. SR-BI is shown in green and nucleus in blue (Hoescht 33342 stain).

Flow Cytometry: SR-BI Antibody [NB400-104] - An intracellular stain was performed on HeLa cells with SR-BI antibody NB400-104AF488 (blue) and a matched isotype control NBP2-24893AF488 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: SR-BI Antibody [NB400-104] - Detection of SR-BI in rat H4IIE total cell lysates and plasma membrane proteins. Photo courtesy of product review by verified customer.

Immunohistochemistry: SR-BI Antibody [NB400-104] - Immunolocalization of SR-BI in adult mink testis using NB400-104. SR-BI labeling is visible at the surface and along the outline of the large vacuole. Photo courtesy of R.M. Pelletier, University of Montreal.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - FFPE tissue section of mouse liver using SR-BI antibody (Lot 8310) at 1:300 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a specific membrane signal of SR-BI protein in the murine hepatocytes.

<b>Genetic Strategies Validation. </b>Western Blot: SR-BI Antibody [NB400-104] - HCV replication was increased by CD36. Silence of CD36 decreased HCV infection and replication (n = 3). Huh7.5 cells were transfected with 0.2 u g CD36-HA plasmid or with 150 pmol siRNA for CD36 (B) for 48 hrs, and then were washed and infected with HCV (150 IU/cell) for 2 hrs, followed by washing and continuously incubating. Intracellular HCV RNA and proteins were detected in 96 hrs, and the cytotoxicity of siRNAs was measured in 96-wells plate with a MTT assay in B. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21808) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - Staining of human adrenal cortex.

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-104] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-SR-BI at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - FFPE tissue section of mouse liver using SR-BI antibody (Lot R-4) at 1:300 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated mainly a membranous signal of SR-BI protein in the murine hepatocytes.

Flow Cytometry: SR-BI Antibody [NB400-104] - Analysis of Huh7 and HepG2 cells using SR-B1 antibody NB400-104. Courtesy of Bruno Sainz, Jr., PhD, University of Illinois at Chicago.

Western Blot: SR-BI Antibody - BSA Free [NB400-104] - Effects of 13-HODE & LA in the presence & absence of PPAR alpha & PPAR gamma selective antagonists on molecular markers of cholesterol homeostasis in RAW264.7 macrophages. RAW264.7 cells were pre-treated without or with the PPAR alpha selective antagonist GW6471 or the PPAR gamma selective antagonist GW9662 & subsequently treated without (vehicle control) or with 2.5 μmol/L 13-HODE or 100 μmol/L LA for 24 h. Afterwards, cells were lysed & subsequently processed for western blotting as described in the materials & methods section. A, Representative immunoblots specific for ABCA1, ABCG1, SR-BI, LXR alpha , & beta -actin which was used for normalization are shown. B, Bars represent data from densitometric analysis & are means ± SD from three independent experiments (n = 3). Data are expressed as percentage of protein concentration of vehicle control cells. Results from statistical analysis are indicated: Significant effects are denoted with superscript letters. Bars marked without a common superscript letter significantly differ (P 2000) (g, h, i, respectively). Bar graph shows the mean percentage of positively stained cells. Error bars depict standard deviations. ****p 0.3) by one-way ANOVA. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29523748), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SR-BI Antibody - BSA Free [NB400-104] - Differentiation & characteristics of iHLCs. (a) Cell morphology & culture conditions at different stages of differentiation from iPSCs into iHLCs. Bright-field images were taken at the indicated time points (scale bar 100 µm). (b) Sequential expression & repression of transcription factors marking successful differentiation. Cells were immunostained with antibodies against Oct 3/4 (pluripotency marker), the endodermal transcription factor GATA-4, & the hepatic marker HNF4 alpha at day 0, 5, & 10 of differentiation (scale bars 20 µm). Shown are representative fluorescence & DIC images. (c) mRNA expression levels of different hepatocyte-specific factors in iHLCs, Huh7.5 cells, & primary human hepatocytes (PHHs) were analysed via qRT-PCR. Expression levels are displayed as fold over iPSCs normalised to GAPDH & 18S rRNA (mean ± SEM, n = 3–4, *p 0.9) by one-way ANOVA. §P < 0.02 by unpaired Student's t-test, relative to 0 h time point. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29523748), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, ChHa, Ha, Md, Pm, Rb, SyHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, ChHa, Ha, Md, Pm, Rb, SyHa

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-104AF350
Alexa Fluor 405	NB400-104AF405
Alexa Fluor 488	NB400-104AF488
Alexa Fluor 532	NB400-104AF532
Alexa Fluor 594	NB400-104AF594
Alexa Fluor 647	NB400-104AF647
Alexa Fluor 700	NB400-104AF700
Alexa Fluor 750	NB400-104AF750
Allophycocyanin	NB400-104APC
Biotin	NB400-104B
CoraFluor 1	NB400-104CL1
DyLight 350	NB400-104UV
DyLight 405	NB400-104V
DyLight 488	NB400-104G
DyLight 550	NB400-104R
DyLight 594	NB400-104DL594
DyLight 650	NB400-104C
DyLight 680	NB400-104FR
DyLight 755	NB400-104IR
FITC	NB400-104F
HRP	NB400-104H
Janelia Fluor 525	NB400-104JF525
Janelia Fluor 549	NB400-104JF549
Janelia Fluor 646	NB400-104JF646
Janelia Fluor 669	NB400-104JF669
PE	NB400-104PE
PerCP	NB400-104PCP
mFluor Violet 450 SE	NB400-104MFV450
mFluor Violet 500 SE	NB400-104MFV500
mFluor Violet 610 SE	NB400-104MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-104SS

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NB100-56708

Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free

(2 Reviews)

(202 Publications)

0.1 mg

NB100-56708

Image of Caspase-3 Antibody (31A1067) - (Pro and Active). Whole cell protein from Jurkat cells treated with and without 2 uM staurosporine as indicated was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 5 ug/ml anti-Caspase 3 in 1% milk, and detected with an anti-mouse HRP secondary antibody using a Femto sensitivity chemiluminescence reagent. Note the detection of both pro-caspase 3 at 35 kDa and the cleaved active caspase 3 at 15-17 kDa.

<b>Biological Strategies Validation. </b>Tissue section of human spleen using 1:200 dilution of Caspase-3 antibody (clone 31A1067). The staining was developed with HRP labeled anti-mouse IgG secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin. This Caspase 3 antibody generated primarily a specific cytoplasmic staining in a subset of spleenocytes with some nuclear signal in a few cells.

Simple Western lane view shows a specific band for Caspase-3 Antibody (31A1067) - (Pro and Active) in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230kDa separation system.

Caspase-3 was detected in immersion fixed paraffin-embedded sections of human bladder tissue using 1:50 dilution of mouse aCaspase-3 Antibody (31A1067) - (Pro and Active) (NB100-56708), for 1 hour at room temperature followed by anti-mouse IgG VisUCyte HRP polymer(VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue).

Paclitaxel in combination with MWE retarded tumor growth in a human bladder carcinoma TSGH 8301 xenograft model. The levels of total (t-PTEN) and phospho-PTEN (p-PTEN) and Caspase 3 in the tumor specimens were determined by Western blotting and then quantified using beta-actin as the protein loading control; the results are expressed as a percentage of the control. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep20417), licensed under a CC-BY license.

An intracellular stain was performed on NIH3T3 cells with Caspase-3 Antibody (31A1067) - (Pro and Active) Antibody NB100-56708AF700 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 700.

<b>Biological Strategies Validation. </b>Lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with VP-16. PVDF membrane was probed with 0.1 ug/mL of mouse monoclonal Caspase-3 Antibody (31A1067) - (Pro and Active) (NB100-56708, Novus Biologicals) followed by 1:2000 dilution donkey anti-mouse IgG.

An intracellular stain was performed on HeLa cells with Caspase-3 Antibody (31A1067) - (Pro and Active) NB100-56708 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems).

Western Blot: Mouse Monoclonal Caspase-3 Antibody (31A1067) - (Pro and Active) [IMGENEX: IMG-144A] [NB100-56708] - Whole cell lysates from MDA-MB-231, MCF-7, SUM149 and SUM159 cells were loaded with 50 ug/lane. 10% SDS-PAGE. Caspase-3 Antibody (NB100-56708) was used for primary antibody: 1:2000, 4℃, overnight. Image from a verified customer review.

Immunocytochemistry/ Immunofluorescence: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - rOPN administration influenced the interaction & balance between Beclin 1 & Caspase‐3 at 24 h after SAH. Double immunofluorescence staining of Caspase‐3 & Beclin 1 in Sham group, SAH + Vehicle group & SAH + rOPN group at 24 h after SAH induction. Sample size is 9, n = 3 per group. Localization of Caspase‐3 can be cytoplasmic & nuclear. Staining in the nucleus is considered to be an indication of active Caspase‐3. The dashed lines & the red box on brain slice images indicate the locations observed. Vehicle, phosphate‐buffered saline; Scale bar = 50 μm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31436915), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - PEM induces YIPF2 upregulation & apoptosis in NSCLC cells.a, b H1792 (a) & H1299 (b) NSCLC cells were treated with PEM at various concentrations (0–10.0 μM) for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2 & ACTB. c Overexpression of YIPF2 in H1299 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, CASP8, CASP3, PARP1 & ACTB. d Knockdown of YIPF2 expression by YIPF2–1 siRNA in A549 NSCLC cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, CASP8, CASP3, PARP1 & ACTB. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32303681), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - Lipofection of top broccoletti-miR candidates does not influence basal & induced apoptosis.(A) BxPc-3 & Bx-Gem cells were transfected as described in Figure 3. Seventy-two hours later, the cells were stained with Annexin V-PE & 7-AAD, followed by FACS analysis. The percentage of Annexin V- & 7-AAD-positive cells is shown. (B) Lipofected BxPc-3 & Bx-Gem cells were stained with an antibody specific for the proliferation marker Ki-67 (green) or the apoptosis marker cleaved fragment of caspase-3 (red), which indicates apoptosis. Representative images at ×100 magnification are shown. The percentage of Ki-67- or caspase-3-positive cells was counted in 18 visual fields, & the means ± SD are shown in the diagram below. (C) BxPc-3 & Bx-Gem were lipofected as described above, & at 24 h later, the cells were treated with gemcitabine (10 nM) or were left untreated. Ninety-six hours after gemcitabine treatment, viability was determined by MTT assay. The data are presented as the means ± SD (**P < 0.01). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32292571), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - PEM induces apoptosis of NSCLC cells via YIPF2-TNFRSF10B axis.a, b A549 (a) & H1792 (b) cells were treated with PEM at 5.0 μM for the indicated times (0, 6, 12, 24, 36 & 48 h). Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B & ACTB. c A549 cells were treated with doxorubicin (DOX) at various concentrations (0–2.0 μM) for 18 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B & ACTB. d Overexpression of YIPF2 in H1792 & H1299 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B & ACTB. e Knockdown of YIPF2 expression by YIPF2–1 siRNA in A549 & H1792 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B & ACTB. f Overexpression of YIPF2 in H1299 cells (left) or Knockdown of YIPF2 expression by YIPF2–1 siRNA in H1792 cells (right) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10A & ACTB. g Three A549 cell lines (Ctrl, YIPF2, YIPF2 + siTNFRSF10B) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B, CASP8, CASP3 & ACTB. h Three H1299 cell lines (Ctrl, siYIPF2–1, siYIPF2–1 + TNFRSF10B (short isoform)) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B, CASP8, CASP3 & ACTB. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32303681), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - Cell numbers positively stained for NeuN, DCX, PCNA & caspase-3 in the subgranular region of the dentate gyrus in WT & RanBP9−/− (KO) mice.(A), DAPI-stained brain sections to show the highlighted subgranular zone within the dentate gyrus region of the hippocampus used for cell counts shown in B. (B), Representative brain sections stained with anti-NeuN, anti-DCX, anti-PCNA, anti-capsase-3 & counter stained with DAPI. Cell counting revealed significantly decreased numbers of NeuN positive cells in RanBP9−/− (KO) brains (22%) compared to WT controls. However DCX, PCNA & caspase positive cell numbers were not significantly altered. In each group, n = 3, data presented as mean± SEM. **, p<0.01 by Student’s t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23840553), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - NFATc1 regulates DcR3 expression at a transcriptional level. (A,B) Immunoblot analysis of whole-cell lysates & quantitative real-time-PCR assaying relative DcR3 mRNA expression of ACHN & 769-P cells 24 h after treatment with cyclosporin A (CsA, 25 μM) or Tacrolimus (FK-506, 50 μM) (A); 48 h post transfection with NFATc1 or an empty vector control (neo) (B). Expression data were normalized to internal 18S rRNA expression (mean ± SEM; n=3; *p<0.05, **p<0.01, ***p<0.001; T-test). (C) Immunoblot analysis of cytoplasmic & nuclear fractions of ACHN & 769-P cells after treatment with LY294002 (50 μM), Everolimus (1 μM), or Cyclosporine A (25 μM). (D) Relative NFATc1–luciferase reporter activity of ACHN & 769-P cells 24 h post transfection with myrAkt or an empty vector control (neo) (mean ± SEM; n=3; *p<0.05, **p<0.01; T-test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24107265), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - rOPN administration elevated the expression of autophagy‐related proteins while suppressing apoptosis in rat brain at 24 h after SAH. The effects of rOPN on expression levels of (A) Beclin 1, mean ± SD is 1.016 ± 0.2262 in Sham group, 2.874 ± 1.147 in SAH + Vehicle group, 4.963 ± 2.05 in SAH + rOPN group, F = 15.52, (B) ATG5, mean ± SD is 0.8908 ± 0.2545 in Sham group, 2.332 ± 0.6431 in SAH + Vehicle group, 4.364 ± 1.309 in SAH + rOPN group, F = 25.02, (C) LC3, mean ± SD is 1 ± 0.1845 in Sham group, 1.755 ± 0.3017 in SAH + Vehicle group, 2.7 ± 0.7957 in SAH + rOPN group, F = 17.23, (D) Cleaved Caspase‐3, mean ± SD is 1.008 ± 0.186 in Sham group, 4.112 ± 1.528 in SAH + Vehicle group, 2.291 ± 0.6268 in SAH + rOPN group, F = 15.86, (E) Bax, mean ± SD is 1.006 ± 0.321 in Sham group, 37.47 ± 10.86 in SAH + Vehicle group, 23.83 ± 8.143 in SAH + rOPN group, F = 33.13, (F) Bcl‐2, mean ± SD is 1.005 ± 0.2736 in Sham group, 0.2309 ± 0.1257 in SAH + Vehicle group, 0.7843 ± 0.2278 in SAH + rOPN group, F = 20.1, in the left hemisphere of rat brain at 24 h after SAH. Sample size is 18, n = 6 per group. Data were presented as mean ± SD. *P < .05, ***P < .001 vs Sham group; #P < .05, ##P < .01 vs SAH + Vehicle group Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31436915), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - RAB8 suppresses PEM-induced apoptosis of NSCLC cells by promoting the removing of TNFRSF10B from plasma membrane to cytoplasm.a, b Knockdown of RAB8 expression by RAB8 siRNA in H1792 (a) & A549 (b) cells in the presence or absence of PEM at 5.0 μM for 36 h. The surface expression of TNFRSF10B was confirmed by flow cytometry analyses. c Overexpression of RAB8 in H1299 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against RAB8, TNFRSF10B, CASP8, CASP3, PARP1 & ACTB. d Knockdown of RAB8 expression by RAB8 siRNA in H1792 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against RAB8, TNFRSF10B, CASP8, CASP3, PARP1 & ACTB. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32303681), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ch, ChHa, Ma

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ch, ChHa, Ma

Applications:

, , , , , , , , , , ,

Applications: , , , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-144A

Alt Catalog# IMG-144A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-56708AF350
Alexa Fluor 405	NB100-56708AF405
Alexa Fluor 488	NB100-56708AF488
Alexa Fluor 532	NB100-56708AF532
Alexa Fluor 594	NB100-56708AF594
Alexa Fluor 647	NB100-56708AF647
Alexa Fluor 700	NB100-56708AF700
Alexa Fluor 750	NB100-56708AF750
Allophycocyanin	NB100-56708APC
Biotin	NB100-56708B
CoraFluor 1	NB100-56708CL1
DyLight 350	NB100-56708UV
DyLight 405	NB100-56708V
DyLight 488	NB100-56708G
DyLight 550	NB100-56708R
DyLight 594	NB100-56708DL594
DyLight 650	NB100-56708C
DyLight 680	NB100-56708FR
DyLight 755	NB100-56708IR
FITC	NB100-56708F
HRP	NB100-56708H
Janelia Fluor 525	NB100-56708JF525
Janelia Fluor 549	NB100-56708JF549
Janelia Fluor 646	NB100-56708JF646
Janelia Fluor 669	NB100-56708JF669
PE	NB100-56708PE
PerCP	NB100-56708PCP
mFluor Violet 450 SE	NB100-56708MFV450
mFluor Violet 500 SE	NB100-56708MFV500
mFluor Violet 610 SE	NB100-56708MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-56708SS
Azide and BSA Free	NBP2-33244

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NB400-132

ABCG1 Antibody - BSA Free

(2 Reviews)

(186 Publications)

0.1 ml

NB400-132

Flow (Intracellular): ABCG1 Antibody - BSA Free [NB400-132] - RAW 246.7 cells were either untreated (A) or serum starved for 24 hours, and then treated with 1uM TO9 for 24 hours (B). An intracellular stain was performed with NB400-132 (blue) and a matched isotype control NB810-56910 (orange). Cells were fixed with 4% paraformaldehyde, following fixation, cells were permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature, followed by Goat Anti-Rabbit Dylight 550-conjugated antibody.

<b>Biological Strategies Validation. </b>Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - Cells were pretreated with 5 uM F12511 or 5 uM MG132 for 30 min and then exposed to 5 or 50 uM 24S-OHC for 6 h. Whole-cell lysates were immunoblotted with antibodies specific for ABCG1 or beta-actin. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31285856/) licensed under a CC-BY license.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - Detection of ABCG1 in J774 mouse macrophages. Image provided by Dr. Angeliki Chroni of the National Centre for Scientific Research Demokritos.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - Raw 264.7 (A), and Raw 264.7 + 10uM T09 18 hr (B) cell lysate using ABCG1 antibody at 2 ug/mL.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - Detection of ABCG1 in mouse peritoneal macrophages. Lane 1: untreated mouse peritoneal macrophages, Lane 2: 22-(R)-hydrocholesterol treated macrophages, Lane 3: T0901713 treated macrophages, Lane 4: T091713 treated macrophages.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - SAA has little impact on the impairment in biliary and fecal excretion of macrophage-derived 3H-cholesterol during endotoxemia. WT and SAAKO mice were injected subcutaneously with PBS or 0.8 ug/g LPS as indicated, followed 4 hr later by intraperitoneal injection of 3H-cholesterol-labeled J774 macrophages. ABCG1 in liver lysates (40 ug protein) was identified by immunoblotting and quantified by densitometry. Results from each of the 3 experiments (n = 4-5 per group per experiment) were expressed relative to PBS-injected WT mice after normalization to beta-actin. A representative western blot is shown. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/jl/2013/283486/), licensed under a CC-BY license.

miR-33 deficiency improved cholesterol efflux in macrophages. A, Quantitative real-time PCR analysis of Abca1 & Abcg1 in macrophages from miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. Values from miR-33+/+Apoe−/− were set at 100%. Values are mean±SE (n=7 each); ***P<0.001. B, Western blotting analysis of ABCA1 & ABCG1 in thioglycollate-elicited peritoneal macrophages from miR-33+/+Apoe−/− & miR-33−/−Apoe−/− mice. GAPDH was used as a loading control. C, Cholesterol efflux from thioglycollate-elicited peritoneal macrophages in the presence or absence of apoA-I (5 or 10 μg/mL). Values are mean±SE (n=6 each); **P<0.01. D, Cholesterol efflux from thioglycollate-elicited peritoneal macrophages in the presence or absence of HDL-C (50 or 100 μg/mL). Values are mean±SE (n=6 each); **P<0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23316322), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

6‐Dihydroparadol increases A) ABCA1 & B) ABCG1, but not C) SR‐BI protein levels in cholesterol‐loaded THP‐1‐derived macrophages. THP‐1 cells were differentiated as described in Figure 2, & then loaded with unlabeled cholesterol for 24 h. Cells were treated with increasing concentrations of 6‐dihydroparadol (1–30 μm) for another 24 h. The protein levels of ABCA1, ABCG1, & SR‐B1 were detected by Western blot analysis. The control was treated with solvent vehicle (0.1% DMSO). As a positive control, TO901317 (5 μm, 24 h) was used. The bar graphs represent mean ± SD from three independent experiments. *p < 0.05, **p < 0.01, & ***p < 0.001 versus control (determined by Student's t‐test or ANOVA with Bonferroni's post hoc test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29802792), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

(A) Chemical structure, (B) cytotoxicity of alpha -asarone, (C) enhancement of cholesterol efflux by alpha -asarone, (D & E) upregulation of ABCA1 & ABCG1 by alpha -asarone & beta -asarone, & (F) elevation of retinoid X receptor (RXR) alpha transcription. J774A.1 murine macrophages were exposed to 50 μg/ml oxidized low-density lipoprotein (LDL) & treated with 1–10 μM purple Perilla frutescens extracts (PPE)-alpha -asarone & 5–10 μM beta -asarone. (B) MTT assay was performed for the measurement of alpha -asarone toxicity. Graph data represent 1 of 4 independent experiments with multiple estimations. Values are expressed as the percentage cell survival relative to the untreated control cells (cell viability, 100%). (C) Cholesterol efflux was expressed as the percentage fluorescence in the medium relative to total fluorescence. (D & E) For the measurement of ABCA1 & ABCG1 expression, total cell lysates were subjected to western blot analysis with a primary antibody against ABCG1 or ABCG1. beta -actin was used as an internal control. Bar graphs (means ± SEM, n=3) represent quantitative densitometric results of the upper bands. Bar graphs denoted without a common letter indicate significant difference, P<0.05. (F) RXR alpha mRNA expression was measured by RT-PCR. GAPDH was used as a housekeeping gene for the co-amplification with RXR alpha . Image collected & cropped by CiteAb from the following publication (//www.spandidos-publications.com/10.3892/ijmm.2015.2101), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

6‐Dihydroparadol increases A) ABCA1 & B) ABCG1 protein stability. THP‐1 cells were differentiated as described in Figure 2. Then cells were loaded with unlabeled cholesterol & treated with 6‐dihydroparadol (10 μm), or solvent vehicle (0.1% DMSO, control) for 24 h. Cells were lysed at different time points (0, 1, 2, 3, 4, & 6 h) after treatment with the protein synthesis inhibitor cycloheximide (CHX, 100 μm). The protein levels of both ABCA1 & ABCG1 were detected by Western blot analysis. The data points represent mean ± SD from three independent experiments. *p < 0.05 & **p < 0.01 versus control at the same time point (determined by Student's t‐test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29802792), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

(A) Upregulation of ABCA1 & ABCG1 & (B) enhancement of cholesterol efflux by purple Perilla frutescens extracts (PPE) in 50 μg/ml Cu2+-oxidized low-density lipoproteins (LDL)-exposed J774A.1 murine macrophages. (A) For the measurement of ABCA1 & ABCG1 expression, total cell lysates were subjected to western blot analysis with a primary antibody against ABCA1 or ABCG1. beta -actin was used as an internal control. Bar graphs (means ± SEM, n=3) represent quantitative densitometric results of the upper bands. (B) Cholesterol efflux was expressed as the percentage of fluorescence in the medium relative to the total fluorescence. Bar graphs denoted without a common letter indicate significant difference, P<0.05. Image collected & cropped by CiteAb from the following publication (//www.spandidos-publications.com/10.3892/ijmm.2015.2101), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - 6‐Dihydroparadol increases A) ABCA1 & B) ABCG1 protein stability. THP‐1 cells were differentiated as described in Figure 2. Then cells were loaded with unlabeled cholesterol & treated with 6‐dihydroparadol (10 μm), or solvent vehicle (0.1% DMSO, control) for 24 h. Cells were lysed at different time points (0, 1, 2, 3, 4, & 6 h) after treatment with the protein synthesis inhibitor cycloheximide (CHX, 100 μm). The protein levels of both ABCA1 & ABCG1 were detected by Western blot analysis. The data points represent mean ± SD from three independent experiments. *p < 0.05 & **p < 0.01 versus control at the same time point (determined by Student's t‐test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29802792), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - Evodiamine enhances apo A1-mediated ChE from THP-1 macrophages & increases ABCA1 protein level. (a) Differentiated THP-1 cells were loaded with [3H]-cholesterol together with the indicated treatments for 24 h. On the next day, the cells were washed twice with PBS & incubated with the same compounds [solvent vehicle control (Veh; ≤0.1% DMSO), evodiamine (1–20 μM), & the PPAR gamma agonist pioglitazone (10 μM) as positive control] with or without 10 µg/mL apo A1. Extracellular as well as intracellular radioactivities were quantified with scintillation counter. Differentiated THP-1-derived macrophages were treated with solvent vehicle control (Veh; ≤0.1% DMSO), evodiamine (10 μM), & the PPAR gamma agonist pioglitazone (10 μM) as positive control. After 24 h incubation, the cells were lysed & 20 μg protein was resolved via SDS-PAGE. Immunodetection was performed with antibodies against the indicated proteins, ABCA1 (b), ABCG1 (c), & SR-B1 (d), & visualized by chemiluminescence detection. All experiments were performed at least three times & data are presented as means ± S.D. vs. solvent vehicle control, *p < 0.05, **p < 0.01, ***p < 0.001, n.s. no significance (ANOVA/Bonferroni). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30038271), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, ChHa, Ha, Pm, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, ChHa, Ha, Pm, Rb

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-132AF350
Alexa Fluor 405	NB400-132AF405
Alexa Fluor 488	NB400-132AF488
Alexa Fluor 532	NB400-132AF532
Alexa Fluor 594	NB400-132AF594
Alexa Fluor 647	NB400-132AF647
Alexa Fluor 700	NB400-132AF700
Alexa Fluor 750	NB400-132AF750
Allophycocyanin	NB400-132APC
Allophycocyanin/Cy7	NB400-132APCCY7
Biotin	NB400-132B
CoraFluor 1	NB400-132CL1
DyLight 350	NB400-132UV
DyLight 405	NB400-132V
DyLight 488	NB400-132G
DyLight 550	NB400-132R
DyLight 594	NB400-132DL594
DyLight 650	NB400-132C
DyLight 680	NB400-132FR
DyLight 755	NB400-132IR
FITC	NB400-132F
HRP	NB400-132H
Janelia Fluor 525	NB400-132JF525
Janelia Fluor 549	NB400-132JF549
Janelia Fluor 646	NB400-132JF646
Janelia Fluor 669	NB400-132JF669
PE	NB400-132PE
PE/Atto594	NB400-132PEATT594
PE/Cy5.5	NB400-132PECY55
PE/Cy7	NB400-132PECY7
PerCP	NB400-132PCP
mFluor Violet 450 SE	NB400-132MFV450
mFluor Violet 500 SE	NB400-132MFV500
mFluor Violet 610 SE	NB400-132MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-132SS

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NBP1-48320

p62/SQSTM1 Antibody - BSA Free

(7 Reviews)

(135 Publications)

0.1 ml

NBP1-48320

<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Cultured HeLa cells were treated with or without 50 uM chloriquine for 24 hours as indicated. Cell lysates were prepared and separated on a 12% gel by SDS-PAGE. Protein was transferred to PVDF membrane and blocked in 5% non-fat milk. The membrane was then probed with 1 ug/ml anti-p62/SQSMT1 in 1% milk and dectected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the upregulation of p62 (arrowhead) in response to chloroquine treatment and the blockage of autophagy.

<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Immunoblotting analysis of LC3 and p62/SQSTM1 expression in thioglycollate-elicited peritoneal macrophages. Macrophages were isolated from C57BL/6 mice fed diets for 12 weeks, treated with or without 100 ng/ml LPS for 18 h with or without 50 nM bafilomycin A1. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S0022227520339523), licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Untreated HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-p628/SQSTM1 at a 1:200 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Vacuolin-1 analogues identified via virtual screening inhibited autophagic flux in HeLa cells. (A) Treatment of HeLa cells with vacuolin-1 analogues (10 M) and BAF (100 nM) failed to further increase the accumulation of both LC3B-II and SQSTM1 as compared to either drug alone. (B) Vacuolin-1 (10 M) or VS6 (10 M) induced the accumulation of yellow LC3B-II puncta in RFP-GFP-LC3B expressing HeLa cells. Scale bar = 10 m. Identification of Novel Vacuolin-1 Analogues as Autophagy Inhibitors by Virtual Drug Screening and Chemical Synthesis. Molecules (2017)

Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Vacuolin-1 analogues identified via virtual screening induced the accumulation of both LC3B-II and SQSTM1 in HeLa cells in a dose dependent manner after a 6 h treatment. Identification of Novel Vacuolin-1 Analogues as Autophagy Inhibitors by Virtual Drug Screening and Chemical Synthesis. Molecules (2017)

Simple Western: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - Drp1+/- mice is protective against autophagy impairment induced by Mn. (A) Representative images of the coronal mouse midbrain section (20um) co-immunostained for DA neurons (TH, red) in the SNpc and GABA neurons (GAD67, green) in the SNpr. Both of these brain regions were removed by laser microdissection for immunoblotting of p62 (top panels) in DA neurons (B) and GABA neurons (C). Total proteins per lane (bottom panels) were used as loading control. (D) Quantified levels of p62 were significantly increased in DA neurons (P = 0.0013), but not in the GABA neurons (P = 0.5457), of the Mn-treated WT mice. Mn did not significantly increase p62 in DA neurons of the Drp1+/- mice (P = 0.8660) No significant baseline level differences between the two genotypes were observed (P = 0.5664 for TH neurons and P = 0.7675 for GABA neurons. Data represent mean +- SEM, n = 5 for WT control, n = 6 for other groups), two-way ANOVA followed by Tukey post-hoc test. (E) Representative confocal images of mitochondrial morphology of DA neurons after TOM20 immunostaining (upper panels), then skeletonized (middle panels) for subsequent analysis. Scale bar 20um. (F, G) Various parameters of mitochondrial morphology were quantified using Fiji MiNA plugin. Data represents mean +- SEM (n = 6 mice per group), analyzed by one-way ANOVA, followed by Tukey post-hoc test Image collected and cropped by CiteAb from the following publication (//molecularneurodegeneration.biomedcentral.com/articles/10.1186/s13024-024-00708-w ), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Autophagy associated protein immunoreactivity in HIV-infected brain tissue. (A) Representative images from five randomly selected fields of cells each examined in duplicate frontal lobe white matter sections for the indicated subject groups. The indicated proteins were labeled red & microglia with the cell-type-specific marker Iba1 (green). Blue staining indicates cell nuclei. Arrow heads indicate examples of higher Iba1 immunoreactivity whereas arrows indicate more focal (punctal) vs. diffuse (filamentous) patterns of autophagy associated protein expression. Scale bar = 10 μm. (B) Quantification of relative Iba1 immunoreactivity from (A). F(3,20) = 6.450, p = 0.0031; ∗p < 0.05 when compared to all other subject groups. Error bars show the SEM for the average values of 2–6 regions from each subject group across the six autophagy associated proteins examined. (C) Quantification of the indicated autophagy associated protein relative immunoreactivity from (A). Beclin 1: F(3,12) = 11.29, p = 0.0008; LC3B: F(3,12) = 1.994, p = 0.1687; APG7/ATG7: F(3,12) = 84.20, p = < 0.0001; ATG5: F(3,12) = 6.218, p = 0.0086; p62/SQSTM1: F(3,12) = 87.04, p = < 0.0001; LAMP1: F(3,12) = 8.317, p = 0.0029. ∗p < 0.05 when compared to HIV-negative; #p < 0.05 when compared to HIV-positive; & Ωp < 0.05 when compared to HIV-positive/NCI subjects. Error bars show the SEM for four regions from each subject group. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/Article/10.3389/fmicb.2015.00653/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - Effects on autophagic activity & dendritic length of neurons exposed to supernatant from HIV-1-infected microglia in combination with morphine. (A) Representative images of neurons with the indicated treatments. Sup, supernatant from uninfected [HIV(-)] & HIV-1-infected [HIV(+)] microglia. Cells were immunolabeled with antibodies to the autophagic activity marker p62/SQSTM1 (red) & the neuronal cell-type-specific marker MAP2 (green). DAPI (blue) staining indicates cell nuclei. (B) Quantification of p62/SQSTM1 immunoreactivity from (A). Data are presented as the percentage of control cells which was set at 100; F(5,24) = 5.882, p = 0.0011; ∗p < 0.05 when compared to HIV(+)Sup + morphine treatment. (C) Western blotting analysis of p62/SQSTM1 & LAMP1 expression levels for the indicated treatments. GAPDH was used as a loading control. Blots are representative of three independent experiments. (D) Measurement of dendrite length from (A). F(5,24) = 26.15, p = < 0.0001; Φp < 0.05 when compared to morphine; Ψp < 0.05 when compared to HIV(-)Sup; Ωp < 0.05 when compared to HIV(-)Sup + morphine; #p < 0.05 when compared to HIV(+)Sup; & ∗p < 0.05 when compared to HIV(+)Sup + morphine treatment. Error bars show the SEM for five randomly selected fields totaling at least 100 cells from each group. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/Article/10.3389/fmicb.2015.00653/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/Immunofluorescence: Rabbit Polyclonal p62/SQSTM1 Antibody [NBP1-48320] - Mice hepatocytes stained with p62/SQSTM1 Antibody. Image from a verified customer review.

Reactivity:

Hu, Mu, Rt, Ch, Po, Bv, Ca, ChHa, In, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Ch, Po, Bv, Ca, ChHa, In, Sh

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-48320AF350
Alexa Fluor 405	NBP1-48320AF405
Alexa Fluor 488	NBP1-48320AF488
Alexa Fluor 532	NBP1-48320AF532
Alexa Fluor 594	NBP1-48320AF594
Alexa Fluor 647	NBP1-48320AF647
Alexa Fluor 700	NBP1-48320AF700
Alexa Fluor 750	NBP1-48320AF750
Allophycocyanin	NBP1-48320APC
Biotin	NBP1-48320B
CoraFluor 1	NBP1-48320CL1
DyLight 350	NBP1-48320UV
DyLight 405	NBP1-48320V
DyLight 488	NBP1-48320G
DyLight 550	NBP1-48320R
DyLight 594	NBP1-48320DL594
DyLight 650	NBP1-48320C
DyLight 680	NBP1-48320FR
DyLight 755	NBP1-48320IR
FITC	NBP1-48320F
HRP	NBP1-48320H
Janelia Fluor 525	NBP1-48320JF525
Janelia Fluor 549	NBP1-48320JF549
Janelia Fluor 646	NBP1-48320JF646
Janelia Fluor 669	NBP1-48320JF669
PE	NBP1-48320PE
PerCP	NBP1-48320PCP
mFluor Violet 450 SE	NBP1-48320MFV450
mFluor Violet 500 SE	NBP1-48320MFV500
mFluor Violet 610 SE	NBP1-48320MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP1-48320SS

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NB110-57130

TRF-2 Antibody - BSA Free

(2 Reviews)

(169 Publications)

0.1 ml

NB110-57130

Western Blot: TRF-2 Antibody [NB110-57130] - Analysis of HeLa whole cell lysate (A), HeLa nuclear cell lysate (B), k562 cell lysate (C), HepG2 cell lysate (D), NIH/3T3 cell lysate (E), CHO cell lysate (F), PC12 cell lysate (G), and Cos7 cell lysate (H) using antibody at a concentration of 2 ug/mL.

Immunohistochemistry-Paraffin: TRF-2 Antibody [NB110-57130] - Analysis in xenografted human breast cancer tissue using DAB with hematoxylin counterstain.

Immunohistochemistry-Paraffin: TRF-2 Antibody [NB110-57130] - Analysis of FFPE human breast cancer tissue with rabbit polyclonal TRF2 antibody at a dilution of 1:200. The staining was developed with HRP-DAB detection method and the counterstaining was performed using hematoxylin. This TRF2 antibody generated an expected nuclear signal in all the cancer cells and the stromal cells. In the tested section, only a subset of myoepithelial cells showed positivity for this protein.

Immunocytochemistry/Immunofluorescence: TRF-2 Antibody [NB110-57130] - NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-TRF-2 Antibody NB110-57130 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: TRF-2 Antibody [NB110-57130] - An intracellular stain was performed on HeLa cells with TRF-2 Antibody NB110-57130 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Chromatin Immunoprecipitation: TRF-2 Antibody [NB110-57130] - Analysis in mouse. Titrated TRF2 antibody to determine concentration required for ChIP experiment. ChIP image submitted by a verified customer review.

<b>Genetic Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: TRF-2 Antibody [NB110-57130] - RNAi-mediated depletion of human separase (ESPL1) induces TIFs. Control scrambled siRNA- (control) and ESPL1 siRNA-treated fibroblasts stained with anti-p53-binding protein 1 (53BP1; green) and anti-TRF2 (red). It is noteworthy that in ESPL1 siRNA-treated cells, 53BP1 signals frequently overlap with TRF2 signals marking the TIFs. Scale bar, 5 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms10405), licensed under a CC-BY license.

Flow Cytometry: TRF-2 Antibody [NB110-57130] - An intracellular stain was performed on HeLa cells with TRF-2 Antibody NB110-57130AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

The DNA damage response at telomeres uncapped through TRF2 depletion does not require BRCA1 or CtIPImmortalized Brca1F/− MEFs were infected with retroviruses expressing the indicated shRNAs and/or Cre recombinase, followed by selection with puromycin for 72 h. Cell extracts were prepared 48 h later & analysed by Western blotting as indicated. SMC1 & tubulin were used as loading controls. *non-specific band.Cells treated as in (A) were fixed 48 h after selection & stained with an anti-53BP1 antibody (green). Telomeres were visualized with a Cy3-conjugated (CCCTAA)3-PNA probe (red). Yellow arrowheads point to 53BP1 foci that co-localize with telomeres.Quantification of TIFs in cells treated as in (B). A minimum of 200 nuclei were scored for each sample. Error bars represent SD of two independent experiments. P-values were calculated using an unpaired two-tailed t-test. *P ≤ 0.05; NS, P > 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25582120), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - Repressive chromatin induced on p21 promoter in TRF2 dependent way. (A–C) Loss of chromatin activation marks H3K4Me, H3K4Me2 in stable TRF2 expressed HT1080 cells (A); increase in LSD1 occupancy in TRF2 over-expressing HT1080 cells. (B) (Full images are shown in Supplementary Figures S6 & S7); Co-immunoprecipitation of TRF2 with LSD1 (immunoprecipitation with anti-TRF2 antibody followed by immunoblotting with anti-LSD1 or anti-TRF2 antibody) (C); (D) Reverse Co-immunoprecipitation of LSD1 with TRF2 (immunoprecipitation with anti-LSD1 antibody followed by immunoblotting with anti-TRF2 antibody); (E) & reduced occupancy of the repressor complex REST, Co-REST & LSD1 on silencing TRF2 on p21 promoter. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-11177-1), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Chromatin Immunoprecipitation: TRF-2 Antibody - BSA Free [NB110-57130] - TRF2 occupancy on G4-motif sites on gene promoters within cells.A, telomeric enrichment was tested for TRF2 ChIP samples performed in HT1080 cells. Telomeric signal was normalized to signal from ALU probe. B, TRF2 occupancy at gene promoter sites in HT1080 cells (endogenous TRF2 & TRF2-overexpressed) quantified using ChIP-qRT PCR. TRF2 ChIP/IgG (mock) enrichment was normalized to 1% input; CTCF promoter (which does not harbor a TRF2 peak) was used as negative control; error bars correspond to S.D., & statistical significance was calculated by paired t test (*, p < 0.05; **, p < 0.01). For the TRF2-overexpressed condition (in red), all p values were <0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31575660), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Chromatin Immunoprecipitation: TRF-2 Antibody - BSA Free [NB110-57130] - Thousands of extra-telomeric TRF2 binding sites found across the genome.A, extra-telomeric TRF2 peaks found in HT1080 cells following TRF2 ChIP-Seq: 20304 TRF2 peaks were common between two independent experiments. B, distribution of common TRF2 peaks around TSSs. Distance from the TSS is shown in kb. C, replicate consistency plot generated using irreproducible discovery rate analysis; 1956 peaks found at ≤0.01 are marked. D, distribution of 1956 TRF2 peaks with IDR 0.01 (TRF2HC peaks) around the TSS. Distance from the TSS is shown in kb. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31575660), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - TRF1HP1 alpha allele-specific protection effects upon si-TRF2-induced telomeric damage. 72 h after transfection, a TRF2 knockdown efficiency with antibody against TRF2 (anti-TRF2) & GAPDH (anti-GAPDH) as loading control. (−) si-non-targeting; (+) si-TRF2. Quantification of TIFs in b si-non-targeting (n = 32–47 nuclei per group) or c si-TRF2. Left *p = 0.0188, right *p = 0.0192, **p = 0.0042, ****p < 0.0001 (n = 31–48 nuclei per group). b, c Significance is assessed by one-way ANOVA & Dunnett’s multiple comparison test with 95% confidence level. Error bars represent s.e.m. Note the similar pattern among TRF1HP1 alpha alleles in c compared to the corresponding allele pattern in Fig. 4b Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30181605), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Chromatin Immunoprecipitation: TRF-2 Antibody - BSA Free [NB110-57130] - TRF2 peaks harbor G-quadruplex motifs genome-wide.A, schematic representation of a G4 motif; sequence pattern with loop/stem & PG4 motif formed by a tetrad of guanine trimers interspersed with loops that can vary in length. B & C, PG4 motifs & TRF2 peaks significantly overlap. High-confidence TRF2-binding sites (TRF2HC peaks) determined by ChIP-Seq in HT1080 cells were significantly enriched in PG4-motif sequences (B), & conversely, PG4-motif sequences were enriched within TRF2HC peaks (C). Nonoverlapping PG4 motifs were considered for analysis; for control analysis, 100 regions of identical length for each TRF2 peak were taken. *, p < 0.05; **, p < 0.01 (Fisher's exact test). Error bars, S.D. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31575660), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - TRF2 transcriptionally regulates p21 expression through promoter occupancy. (A) Quantitative ChIP using TRF2 antibody gives enriched binding of TRF2 on p21 promoter in HT1080 & MDA-MB-231 cells, IgG was used as isotypic antibody control; normalized with 10% input (data represented as mean ± SEM, for three replicates). (B,C) TRF2 represses p21 promoter activity. In luciferase assay, si-RNA-mediated silencing or TRF2 over expression resulted in increased (B, *p value < 0.05, Student’s t-test; data represented as mean ± SEM of three replicates) or reduction (C, *p value < 0.05, Student’s t-test; data represented as mean ± SEM of three replicates) in p21 promoter activity in HT1080 & MDA-MB-231 cells, respectively; over expression of TRF2 devoid of DNA binding [deletion of basic (delB), myb (delM) & both basic/myb (delB-delM) domains] resulted in partial or complete rescue of p21 promoter activity in HT1080 & MDA-MB-231 cells (data represented as mean ± SEM, for three replicates) (C). (D,E) HT1080 cells over expressing TRF2 show reduced p21 protein expression, bar graph shows the densitometry analysis of three different immunoblot replicates (*p value < 0.05, Student’s t-test; data represented as mean ± SEM of three replicates). (E) (Full images are shown in Supplementary Figure S5); TRF2 silencing results in increase in p21 protein, bar graph shows the densitometry analysis of three different immunoblots (*p value < 0.05, Student’s t-test; data represented as mean ± SEM of three replicates) (F); & mRNA expression in HT1080 cells (*p value < 0.05, Student’s t-test; data represented as mean ± SEM of three replicates, GAPDH used as internal control for real-time PCR). Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-11177-1), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Chromatin Immunoprecipitation: TRF-2 Antibody - BSA Free [NB110-57130] - TRF2 occupancy at gene promoters is sensitive to intracellular G-quadruplex–binding ligand 360A.A, TRF2 occupancy at gene promoter sites was checked by ChIP qRT-PCR in HT1080 cells following overexpression of TRF2 in the presence or absence of 360A. Error bars, S.D. from three independent experiments; CTCF promoter was used as a negative control. B, TRF2 level was checked by Western blotting in the presence of the ligand 360A in both untransfected & TRF2 transient overexpression conditions. Overexpression was also confirmed by probing for DDK tag. GAPDH was used as loading control. C, expression of target genes was analyzed by qRT-PCR in HT1080 cells following overexpression of TRF2 in the presence or absence of 360A. GAPDH expression was used for normalization; error bars, S.D. from three independent experiments. Statistical significance was calculated by paired t test (*, p < 0.05; **, p < 0.01). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31575660), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - A. Co-IP analysis of the interaction between DSP & TRF2 in HEK293 cells & MCF-7 cells. B. Co-IP analysis of the interaction between DSP & TRF2 in DSP-EGFP transfected HEK293 cells or DSP△NLS-EGFP transfected HEK293 cells. C. DSP pull down by telomere targeting CASID with the condition of TRF2 knockdown in HEK293 cells. Histone H3 is the loading control. D.TRF1 & TRF2 pull-down assay by telomere targeting CASID with the condition of DSP knockdown in HEK293 cells. E. Left, Dot blot of telomere DNA repeats in chromatin pull down by DSP with the condition of TRF1 or TRF2 knockdown. The dot-blot was performed at least three times & the represented one is shown. Right, the histogram shows the quantitated results of the dot blots. The triple asterisks (***) indicates a p-value less than 0.01. ns. indicates the insignificance difference. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31595153), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody - BSA Free [NB110-57130] - TERRA & telomere transcription in ALT cells.(a) TERRA northern blot hybridizations of RNA from the indicated cell lines (VA13: WI-38 VA13; 1.2.11: HeLa 1.2.11) pre-treated with RNaseA or left untreated. Ethidium bromide stained 18S ribosomal RNA (rRNA) is shown to control for loading. Long TERRA molecules comprised between the wells of the gels (w) & 28S rRNA are indicated. (b) TERRA CpG-island promoter methylation analysis of the indicated cell lines. Genomic DNA was digested with the methylation sensitive restriction enzyme MspI or its methylation insensitive isoschizomer HpaII. DNA was hybridized using a radioactively labelled probe detecting TERRA promoter CpG-island repeats. Nomet: fragments corresponding to unmethylated restriction products. (c) Dot blot hybridization of DNA immunoprecipitated with antibodies against phosphorylated Serines S2 & S5 of RNA polymerase II C-terminal domain. Hybridizations were performed with a telomeric probe. Quantifications are shown at the bottom. (d) Bars & error bars are averages & s.d. from three independent experiments. (e) Examples of TERRA FISH in the indicated cells. TERRA is shown in red, DAPI-stained DNA in blue. Scale bar, 9 μm. (f) IF/FISH experiments in the indicated cell lines. TERRA is in red, TRF2 in green & PML in blue. In the merge panels, arrowheads point to nuclear foci where the three factors co-localize. Scale bar, 9 μm. (g) Information surface at 0.01 μm detail level of three TERRA-containing APBs. TERRA is in red, TRF2 in green & PML in cyan. Images were generated with Three-Dimensional Structured Illumination Microscopy (3D-SIM). Scale bars, 0.4 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25330849), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - TRF2 occupancy at gene promoters is sensitive to intracellular G-quadruplex–binding ligand 360A.A, TRF2 occupancy at gene promoter sites was checked by ChIP qRT-PCR in HT1080 cells following overexpression of TRF2 in the presence or absence of 360A. Error bars, S.D. from three independent experiments; CTCF promoter was used as a negative control. B, TRF2 level was checked by Western blotting in the presence of the ligand 360A in both untransfected & TRF2 transient overexpression conditions. Overexpression was also confirmed by probing for DDK tag. GAPDH was used as loading control. C, expression of target genes was analyzed by qRT-PCR in HT1080 cells following overexpression of TRF2 in the presence or absence of 360A. GAPDH expression was used for normalization; error bars, S.D. from three independent experiments. Statistical significance was calculated by paired t test (*, p < 0.05; **, p < 0.01). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31575660), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - A. Gene ontology analysis of proteins identified by CASID targeting telomeres. B. Western blot detection of TRF1, POT1, & DSP in the proteins pulled down by CASID with the presence of telo-sgRNA in HEK293 cells. Input represents the initial nuclear extract before the pull-down mediated by CASID. C. left, ChIP assay of telomere binding of TRF2 or DSP in MCF-7 cells & HEK293 cells. The chromatin DNA precipitated by anti-DSP & anti-TRF2 antibodies were analyzed by dot blot using the probes recognizing telomere (Telo-p) or Alu DNA repeats (Alu-p). The detection of Alu DNA repeats serves as negative control. The dot-blot was performed at least three times & the represented one is shown. Right, the histogram shows the quantitated results of the dot blots. The triple asterisks (***) indicates a p-value less than 0.01. D. Fluorescence imaging of DSP-EGFP in MCF-7 cells & U2OS cells. The EGFP signal indicates the localization of DSP-EGFP (green) & the Telo-FISH signal indicates the localization of telomeres (red). The overlapping between DSP-EGFP & Telo-FISH is in yellow as revealed in the enlarged nuclear area. The dCAS9-EGFP construct without telo-sgRNA was used for U2OS transfection & the imaging of EGFP serves as negative control. E. DSP localization in interphase & pro-metaphase nucleus. DSP-EGFP indicates the DSP locations & DAPI indicates the nucleus. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31595153), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - TRF2 interacts with lamin A/C.(a) Pull down of endogenous TRF2 in IMR90s. The top panel shows a blot probed for endogenous lamin A/C, & the bottom panel shows a blot with the same samples probed for TRF2. (b) Pull down of endogenously expressed lamin A/C in IMR90s. The top panel shows a blot probed for endogenous TRF2, & the bottom panel shows a blot with the same samples probed for endogenous lamin A/C. (c) Pull down of endogenous lamin A/C in IMR90s exogenously expressing GFP, GFP-TRF2, GFP-TRF delta B delta M or GFP-TRF1. The top panel shows a blot probed for GFP, & the bottom panel shows the same blot probed for lamin A/C. (d) Pull down of endogenous TRF2 in IMR90s exogenously expressing GFP, GFP-LMNA or progerin (GFP-LMNA delta 50). The top panel shows a blot probed for GFP, & the bottom panel shows the same blot probed for TRF2. Asterisks denote IgG bands. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25399868), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - Widespread dysregulation of shelterin protein expression in bortezomib-treated HEL & BGC-823 cellsA. & B. mRNA levels of shelterin factors TRF1, TRF2, TPP1, POT1, RAP1 & TIN2 in bortezomib-treated cells. Cells were treated with bortezomib for 24 hours & qPCR was used for quantitative assays. The levels of each target mRNA in bortezomib-treated cells were expressed as percentages of those in untreated cells. (A) HEL cells & (B) BGC-823 cells. C. & D. Immunoblotting assessment of TRF1, TRF2 & POT1 protein expression in bortezomib-treated cells. Same sets of cells above were analyzed for TRF1, TRF2 & POT1 protein levels & shown was representative of three independent experiments. (C) HEL cells & (D) BGC-823 cells. * & **: P < 0.05 & 0.01, respectively. BTZ, bortezomib. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.5752), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - Telomere deprotection contributes to replication stress lethality. a Western blots of whole cell extracts from HT1080 6TG cells stably transduced with control, TRF2 shRNA (TRF sh-F) or TRF2 over expression (TRF2OE) vectors. b Representative images of cyto-centrifuged chromosome spreads stained with DAPI (blue), gamma -H2AX IF (red) & telomere PNA (green) from control, TRF sh-F or TRF2OE HT1080 6TG cells treated with DMSO or APH. Scale bar represents 10 µm. c, d Quantitation of mitotic telomere DDR foci from Control sh & TRF2 sh-F cells (c) or vector & TRF2OE cells (d) ± DMSO or APH (three biological replicates scoring n = 50 mitotic spreads per replicate compiled in a Tukey box plot, Mann–Whitney test). e Difference in the mean number of mitotic telomeric gamma -H2AX foci between HT1080 6TG TRF2 sh-F or TRF2OE cells & their appropriate vector control. These are a different representation of the same data shown in (c, d) (mean ± s.e.m., n = 3 three biological replications, Student’s t-test). f Mitotic duration to cell death in APH treated control, TRF2 sh-F & or TRF2OE cells (three biological replicates scoring ≥267 mitotic death events per condition are shown in a dot plot, mean ± s.e.m., Mann–Whitney test). g Mitotic outcome of control, TRF2 sh-F & TRF2OE cells treated with APH or DMSO (mean ± s.e.m., n = 3 biological replicates of at least 267 mitoses per condition, Fisher’s Exact Test). For all panels, *p < 0.05, **p < 0.01. Source data are provided as a Source Data file Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31530811), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - A. TRF2 or DSP1 pull down by telomere targeting CASID with the condition of MST-312 treatment (left) or hTERT knockdown (right) in MDA-MB-231cells. B. Southern blot of telomere DNA from MDA-MB-231cells treated with MST-312 or with hTERT knockdown. C. Top, Dot blot of telomere DNA repeats from the chromatin pull down by DSP from the cells treated by MST-312 (left) or hTERT knockdown (right). Alu DNA repeats serve as negative control. Bottom, the histogram shows the quantitated results of the dot blots. The triple asterisks (***) indicates a p-value less than 0.01. D. Fluorescence image of DSP-EGFP transfected MDA-MB-231 cells with the conditions of MST-312 treatment or hTERT knockdown. E. Pull down assay of DSP by CASID targeting telomere in WI38 cells with or without ectopically expression of hTERT gene. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31595153), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TRF-2 Antibody - BSA Free [NB110-57130] - Attenuation of bortezomib-induced shelterin protein dysregulation by hTERT over-expressionA. & B. Cells expressing ectopic hTERT were treated with bortezomib for 24 hours & mRNA levels of shelterin proteins then analyzed using qPCR. The levels of each target mRNA in bortezomib-treated cells were expressed as percentages of those in untreated cells. (A) HEL-hTERT cells & (B) BGC-823-hTERT cells. C. & D. Immunoblotting assessment of TRF1, TRF2 & POT1 protein expression in bortezomib-treated cells. Same sets of cells above were analyzed for TRF1, TRF2 & POT1 protein levels & shown was representative of three independent experiments. (C) HEL-hTERT cells & (D) BGC-823-hTERT cells.* & **: P < 0.05 & 0.01, respectively. BTZ, bortezomib. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.5752), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody - BSA Free [NB110-57130] - RNaseH1 depletion leads to RPA activation at ALT telomeres.(a) HeLa & U2OS cells were transfected with the indicated siRNAs & 48 & 72 h later protein extracts were prepared. Western blot analysis was performed using antibodies against RNaseH1, RPA32 phosphorylated at Serine 33 (pSer33), total RPA32 & KAP1 (loading control). The asterisk indicates a cross-reacting band. Cells treated for 6 h with 5 mM hydroxyurea (HU) were used as controls for pSer33 activation. (b) SiRNA transfected cells were subjected to indirect immunofluorescence using antibodies against TRF2 (to visualize telomeres; red) & pSer33 (green). DNA was counterstained with DAPI (blue). Arrows point to examples of pSer33 foci co-localizing with TRF2 (TIFs). Scale bar, 9 μm. Cells treated for 6 h with 5 mM HU were used as controls for pSer33 activation. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25330849), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, ChHa, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, ChHa, Pm

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB110-57130AF350
Alexa Fluor 405	NB110-57130AF405
Alexa Fluor 488	NB110-57130AF488
Alexa Fluor 532	NB110-57130AF532
Alexa Fluor 594	NB110-57130AF594
Alexa Fluor 647	NB110-57130AF647
Alexa Fluor 700	NB110-57130AF700
Alexa Fluor 750	NB110-57130AF750
Allophycocyanin	NB110-57130APC
Biotin	NB110-57130B
CoraFluor 1	NB110-57130CL1
DyLight 350	NB110-57130UV
DyLight 405	NB110-57130V
DyLight 488	NB110-57130G
DyLight 550	NB110-57130R
DyLight 594	NB110-57130DL594
DyLight 650	NB110-57130C
DyLight 680	NB110-57130FR
DyLight 755	NB110-57130IR
FITC	NB110-57130F
HRP	NB110-57130H
Janelia Fluor 525	NB110-57130JF525
Janelia Fluor 549	NB110-57130JF549
Janelia Fluor 646	NB110-57130JF646
Janelia Fluor 669	NB110-57130JF669
PE	NB110-57130PE
PerCP	NB110-57130PCP
mFluor Violet 450 SE	NB110-57130MFV450
mFluor Violet 500 SE	NB110-57130MFV500
mFluor Violet 610 SE	NB110-57130MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB110-57130SS

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NB100-690

alpha Tubulin Antibody (DM1A) - BSA Free

(14 Reviews)

(166 Publications)

0.1 ml

NB100-690

Immunohistochemistry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of colon tissue. Sections were formalin fixed and embedded with paraffin. Sodium citrate heat mediated antigen retrieval for 20 min. Incubated with primary antibody for 15 min at a 5 ug/ml concentration. Corner image is staining with secondary only.

Immunohistochemistry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of formalin fixed paraffin embedded heart sections. Used at a dilution of 1:500.

Western Blot: alpha Tubulin Antibody (DM1A) [NB100-690] - Analsis of alpha tubulin in 9 cell lysates. Lane 1. HeLa; Lane 2. JURKAT; Lane 3. COS7; Lane 4. NIH-3T3; Lane 5. PC-12; Lane 6. RAT2; Lane 7. CHO; Lane 8. MDBK; Lane 9. MDCK

Flow Cytometry: alpha Tubulin Antibody (DM1A) [NB100-690] - Intracellular flow cytometric staining of 1 x 10^6 CHO (A) and HEK-293 (B) cells using alpha Tubulin antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1x10^6 cells was used.

Immunomicroscopy: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of HeLa cells, green staining is alpha tubulin whereas red is DNA stained with propidium iodide.

Western Blot: alpha Tubulin Antibody (DM1A) [NB100-690] - GS treatment increases markers of beiging in 3T3-L1 adipocytes. GS treatment upregulates markers of beiging, including beta-3AR (C) proteins. Data presented as mean +/- SEM from n = 4 replicates per group. * p 0 for sensitive & <0 resistant. (D) Apoptotic H1299 (null p53), A549 (wild type p53), Bm7-shGFP (mutant p53), & Bm7-shTP53 (knock-down p53) cells treated with simvastatin were detected using flow cytometry, *P < 0.05 & **P < 0.01. (E) Apoptotic HCC827-shGFP (mutant p53) & HCC827-shTP53 (knock-down p53) cells treated with simvastatin & cisplatin were detected using flow cytometry, *P < 0.05 & **P < 0.01. (F) Western blots of indicated proteins involved in apoptosis & autophagy in both Bm7 & HCC827 cells with control (shGFP) & p53 knockdown (shTP53) treated with simvastatin is shown. MDM2, murine double minute 2; AKT, serine–threonine kinase; PARP, poly (ADP-ribose) polymerase; mTOR, mammalian target of rapamycin; WT, wild type. Full-length blots/gels are presented in Supplementary Fig. 1. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31892709), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: alpha Tubulin Antibody (DM1A) - BSA Free [NB100-690] - Pre-treatment with 0.5 mM sodium arsenite (SA) enhances permissivity in a cell-type-specific manner across reovirus strains. (A) CV-1, HeLa, L929, or HPDE cells were left untreated (no SA) or were treated with 0.5 mM SA for 30 min prior to infection (Pre-SA). Following this, cells were infected with T3D such that ~20% to 50% of cells were infected & at 18 h p.i. cells were fixed & immunostained for μNS & DAPI to visualize viral factories (VFs). The percent of cells containing VFs was quantified ((# of cells containing VFs/total # of cells) × 100) from three independent experiments. The expression level of μNS (B) & μ1 (C) was determined in CV-1, L929, or HeLa cells either left untreated (no SA) or treated with 0.5 mM SA for 30 min (Pre-SA) before infection with T3D at MOI = 1. At 18 h p.i., cells were harvested & the expression level of the indicated proteins was determined by immunoblot. M = mock. Densitometry analysis of the band intensity for μNS & μ1 was adjusted to the matched alpha -tubulin loading control for two independent experiments. Columns represent mean ± SEM. (D) CV-1; (E) L929; or (F) HeLa cells were left untreated (no SA) or were treated with 0.5 mM SA prior to infection (Pre-SA). Cells were then infected with the reovirus strains, T3D, T1L, or T3A, as described in (A). At 18 h p.i., cells were fixed & immunostained for μNS & DAPI to detect VFs. The percent of cells containing VFs was quantified ((# of cells containing VFs/total # of cells) × 100) from at least two independent experiments. * p < 0.05; ** p < 0.01; two-tailed unpaired t test. The error bars indicate S.D. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31216693), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Alpha Tubulin (DM1A) was detected in immersion fixed U-2 OS human osteosarcoma cell line using Mouse anti-Alpha Tubulin (DM1A) Protein G Purified Monoclonal Antibody conjugated to Alexa Fluor ® 488 (Catalog # NB100-690AF488) (green) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (dark blue). Cells were imaged using a 100X objective and digitally deconvolved.

An intracellular stain was performed on U-2 OS human osteosarcoma cell line with Mouse anti-alpha Tubulin (DM1A) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor ® 488 (Catalog # NB100-690AF488, blue histogram) or matched control antibody (orange histogram) at 5 µg/mL for 30 minutes at RT.

An intracellular stain was performed on A431 human skin carcinoma cell line using Mouse anti-alpha Tubulin (DM1A) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor ® 647 (Catalog # NB100-690AF647, blue histogram) or matched control antibody (orange histogram) at 2.5 µg/mL for 30 minutes at RT.

Alpha Tubulin (DM1A) was detected in immersion fixed NIH-3T3 Mouse fibroblast cell line using Mouse anti-Alpha Tubulin (DM1A) Protein G Purified Monoclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB100-690AF488) (green) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (dark blue). Cells were imaged using a 100X objective and digitally deconvolved.

Reactivity:

Hu, Mu, Rt, Po, Av, Bv, Ca, Ch, ChHa, Dr, Fu, Gp, Gt, Ha, Pa, Pm, Pm, Rb, Xp, Ye

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Av, Bv, Ca, Ch, ChHa, Dr, Fu, Gp, Gt, Ha, Pa, Pm, Pm, Rb, Xp, Ye

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-690AF350
Alexa Fluor 405	NB100-690AF405
Alexa Fluor 488	NB100-690AF488
Alexa Fluor 532	NB100-690AF532
Alexa Fluor 594	NB100-690AF594
Alexa Fluor 647	NB100-690AF647
Alexa Fluor 700	NB100-690AF700
Alexa Fluor 750	NB100-690AF750
Allophycocyanin	NB100-690APC
Biotin	NB100-690B
CoraFluor 1	NB100-690CL1
DyLight 350	NB100-690UV
DyLight 405	NB100-690V
DyLight 488	NB100-690G
DyLight 550	NB100-690R
DyLight 594	NB100-690DL594
DyLight 650	NB100-690C
DyLight 680	NB100-690FR
DyLight 755	NB100-690IR
FITC	NB100-690F
HRP	NB100-690H
Janelia Fluor 525	NB100-690JF525
Janelia Fluor 549	NB100-690JF549
Janelia Fluor 646	NB100-690JF646
Janelia Fluor 669	NB100-690JF669
PE	NB100-690PE
PerCP	NB100-690PCP
mFluor Violet 450 SE	NB100-690MFV450
mFluor Violet 500 SE	NB100-690MFV500
mFluor Violet 610 SE	NB100-690MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP3-38646SS
BSA Free	NBP3-38646
BSA Free	NB100-690SS
Azide and BSA Free	NBP2-80570

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NB100-56566

TLR4 Antibody (76B357.1) - BSA Free

(5 Reviews)

(148 Publications)

0.1 mg

NB100-56566

Immunohistochemistry: TLR4 Antibody (76B357.1) [NB100-56566] - Pericryptal Myofibroblasts are Responsible for Increased TLR4 Expression in a Subset of CRCs. Double-stained immunofluorescence for TLR4 (green) and vimentin (red) in normal (I), adenoma (II), and colon adenocarcinoma (III) (10x). In the stromal compartment of CRCs, immunofluorescent staining for TLR4 localized to the pericryptal myofibroblasts in a subset of samples. Image collected and cropped by CiteAb from the following publication (//www.jeccr.com/content/33/1/45), licensed under a CC-BY license.

Western Blot: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using 2 ug/mL on (A) human intestine and 6 ug/mL on (B) mouse intestine and (C) rat intestine lysate.

Flow (Intracellular): TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using PE conjugate of NBP2-27149. An intracellular stain was performed on Jurkat cells with TLR4 antibody (76B357.1) NBP2-27149PE (blue) and an isotype control MAB004 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.

Immunocytochemistry/Immunofluorescence: TLR4 Antibody (76B357.1) [NB100-56566] - RH-30 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-TLR4 Antibody (76B357.1) NB100-56566 at 1 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunohistochemistry: TLR4 Antibody (76B357.1) [NB100-56566] - Immunofluorescent staining of TMAs. Representative tissue cores from normal (I), adenomatous polyps (II), and CRC (III and IV) are shown. Image collected and cropped by CiteAb from the following publication (//www.jeccr.com/content/33/1/45), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of rat salivary gland tissue section at 1:100 dilution. The antibody generated a membrane-cytoplasmic staining in the tissue with stronger signal in ductal epithelial cells.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Tissue section of normal human skin stained with antibody at 5 ug/mL. Membrane-cytoplasmic immunopositivity of TLR4 was primarily observed in the pigmented basel cells and the adjacent keratinocytes in the epidermal layer.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on RH30 cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human skin stained with at 5 ug/mL, peroxidase-conjugate and DAB chromogen. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of TLR4 in FFPE human colon tissue using an isotype control (top) and NB100-56566 (bottom) at 5 ug/mL.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human testis tissue stained with antibody at 5 ug/mL.

Immunohistochemistry-Frozen: TLR4 Antibody (76B357.1) [NB100-56566] - This image is TLR4(green) and nucleus(blue) at area postrema of the adult male mouse brain, x20 magnification. Primary antibody diluted 1:500. IHC-Fr image submitted by a verified customer review.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of formaldehyde fixed THP-1 cells (human monocytic leukemia cells) using 2 ug/10^6 cells TLR4 antibody (clone 76B357.1) with detection employing a donkey anti-mouse IgG (H+L) cross adsorbed secondary antibody, (DyLight 488 conjugated). Isotype control samples incubated with mouse IgG2b isotype control antibody were processed in parallal under the same assay conditions.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on Jurkat cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on Raw264.7 cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Representative images of immunohistochemical staining of pancreatic adenocarcinoma upregulated factor (PAUF) and toll-like receptor 4 (TLR4) in human ovarian cancer specimens. The top row (A,B) represents normal ovarian tissue, middle row (C,D) shows immunoreactivity of ovarian carcinomas and the bottom row (E,F) indicates immunonegativity of ovarian caricinoma. The stromal cells of normal ovarian tissue show immunonegativity for PAUF and TLR4. The scale bar represents 100 μm.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Increased toll‐like receptor 4 (TLR4) expression in the myocardium of chronic heart failure (CHF) rats. (A) TLR4 mRNA levels in infarct & remote myocardium of sham & CHF rats (n = 6/group). (B) Representative Western blot images & (C) quantification of TLR4 proteins in infarct & remote myocardium of sham & CHF rats (n = 4/group). (D) Representative immunohistochemistry images of heart sections stained with TLR4 (green) & CD45 (red). The yellow box indicates the enlarged area shown on the right (data are means ± SD, *P < 0.05, **P < 0.01 versus respective sham). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26290459), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Regulation of FOXC1 on TLR3/4 expression in mice subjected to myocardial ischaemia (MI). The siRNA against FOXC1 (si‐FOXC1) or FOXC1 adenovirus (Ad‐FOXC1) was injected into the left ventricle just after LAD ligation to generate FOXC1 knockdown (A) or FOXC1 overexpression (B), & the negative control (NC) siRNA & Ad‐GFP served as control, respectively. The mRNA (upper panel) & protein (middle & lower panel) levels of TLR3/4 were determined after 2 wk. Data are means ± SEM aP < .05, AP < .01 vs respective MI Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31517441), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Representative images of immunohistochemical staining of pancreatic adenocarcinoma upregulated factor (PAUF) & toll-like receptor 4 (TLR4) in human ovarian cancer specimens. The top row (A,B) represents normal ovarian tissue, middle row (C,D) shows immunoreactivity of ovarian carcinomas & the bottom row (E,F) indicates immunonegativity of ovarian caricinoma. The stromal cells of normal ovarian tissue show immunonegativity for PAUF & TLR4. The scale bar represents 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30111860), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Increased toll‐like receptor 4 (TLR4) expression in the surviving cardiomyocytes of chronic heart failure (CHF) rats. (A) Representative immunofluorescent images of TLR4 in cardiomyocytes isolated from sham & CHF rats. (B) TLR4 mRNA levels in cardiomyocytes isolated from sham & CHF rats. (C) Representative Western blot images & (D) quantification of TLR4 proteins in cardiomyocytes isolated from sham & CHF rats (data are means ± SD, n = 6/group, **P < 0.01 versus sham). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26290459), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Up‐regulation of FOXC1 & TLR3/4 protein levels under myocardial ischaemia. Representative Western blot images & quantitative data for FOXC1, TLR3 & TLR4 proteins in ischaemia models of mice (A), H9c2 cells (B) & NRVMs (C) are shown herein. Validation data of the antibodies for TLR3 & TLR4 are shown in Figure S1. Data are means ± SEM. n = ~4‐5/group. P values from the one‐way ANOVAs: 0.018 (TLR3 protein in mice), .002 (TLR4 protein in mice) & 0.006 (FOXC1 protein in mice). aP < .05, AP < .01 vs control Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31517441), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - (a) Cell viability of MM cells exposed to increasing doses of LPS for 24 h. (b) Immunoblotting analysis of TLR4 protein expression levels in H929 & L363 cell lines after incubation with various LPS concentrations. Probing with beta -ACTIN was used as total protein loading reference. (c) Flow cytometry analysis for TLR4 expression of MM cell lines before & after exposure to 1 μg/ml LPS for 24 h. (d) Cell viability of MM cell lines pre-treated with 0.5 μg/ml TLR4 inhibitor for 24 h before stimulation with LPS for 24 h. (e) Cell vViability of H929 & JJN3 cell lines exposed to increasing concentrations of TLR4 inhibitor for 48 h. beta -ACTIN probing was used as reference for total protein input. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30824741), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - TLR4 mRNA (a1) & protein (a2) expression in MM cell lines. (b) TLR4 mRNA expression of 16 CD138+ selected MM patients as determined by PCR & agarose gel electrophoresis. Probing with beta -ACTIN was used as total protein loading reference, whereas beta -ACTIN gene expression was used as reference for RNA input. beta -ACTIN probing & beta -ACTIN mRNA expression were used as reference for total protein & mRNA input, respectively. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30824741), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Regulation of FOXC1 on TLR expression. H9c2 cells were transfected with adenovirus or siRNA to overexpress or knock down FoxC1, with GFP adenovirus (Ad‐GFP) & negative control (NC) siRNA serving as control, respectively. A, FoxC1 overexpression increased Tlr3/4 mRNA & protein levels, under both control & ischaemic conditions. P values from the one‐way ANOVAs: <.001 (TLR3 mRNA), .002 (TLR4 mRNA), <.001 (FOXC1 mRNA), <.001 (TLR3 protein), <0.001 (TLR4 protein) & <.001 (FOXC1 protein). B, FoxC1 knockdown decreased Tlr3/4 mRNA & protein levels. P values from the one‐way ANOVAs: .001 (TLR3 mRNA), <.001 (TLR4 mRNA), <.001 (FOXC1 mRNA), .002 (TLR3 protein), <.001 (TLR4 protein) & <.001 (FOXC1 protein). C, FoxC1 overexpression up‐regulated the mRNA expression of multiple Tlr subtypes. P values from the one‐way ANOVAs: .022 (TLR1 mRNA), .047 (TLR2 mRNA), .002 (TLR3 mRNA), .013 (TLR4 mRNA), .039 (TLR5 mRNA), .003 (TLR6 mRNA), .203 (TLR7 mRNA), .078 (TLR8 mRNA) & .009 (TLR9 mRNA). Data are means ± SEM of 4 independent experiments. aP < .05, AP < .01 vs. control; bP < .05, BP < 0.01 vs. ischaemia Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31517441), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Immunofluorescent staining of TMAs. A) Low power (10x) view of NCI TMA slide stained for TLR4 (green), intestinal epithelium/pan-cytokeratin (red), & nucleus/DAPI (blue). B) Representative tissue cores from normal (I), adenomatous polyps (II), & CRC (III & IV) are shown. C & D) TLR4 staining score by tissue type & tissue compartment (stroma vs epithelium) are shown. C) TLR4 staining in the tumor stroma had a significantly higher average intensity score for stages 3 & 4 CRC when compared to stage 1. D) TLR4 staining in the tumor epithelium had a significantly higher average intensity score for stages 2 & 3 when compared to stage 1. E) TLR4 staining by compartment broken down by stage (controlling for grade) & grade (controlling for stage). Image collected & cropped by CiteAb from the following publication (//jeccr.biomedcentral.com/articles/10.1186/1756-9966-33-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Effect of Cucumis sativus L. extract (CSE) on LPS induced TLR4 expression. a Expression of TLR4 mRNA in pAECs treated with LPS (10 μg/ml) for different time (1, 7, 24 h) in the presence or absence of increasing doses of CSE (0.02; 0.2; 2 mg/ml). mRNA expression of TLR4 is determined by quantitative PCR; relative expression was calculated as fold of change in respect to the control cells & error bar represents the range of relative expression. Different letters above the bars indicate significant differences. b Representative Western Blot of TLR4 & relative housekeeping alpha -tubulin were reported. c Expression of TLR4 protein in pAECs treated with LPS (10 μg/ml) in the presence or absence of increasing doses CSE (0.02; 0.2; 2 mg/ml); data shown are representative of at least three independent experiments & represent the mean ± SEM. Different letters above the bars indicate significant differences (p < 0.05 ANOVA post hoc Tukey’s test) Image collected & cropped by CiteAb from the following publication (//bmccomplementalternmed.biomedcentral.com/articles/10.1186/s12906-018-2254-1), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Activation receptors are shed during intrinsic repair. (A) BMDM were unchallenged (control) or challenged with 500 HU/mL SLO WT, PFO or an equivalent mass of SLO ML for 10 min at 37 °C & centrifuged at 2000 × g for 5 min to yield cell pellet (C). Supernatants were spun at 100,000xg for 40 min at 4 °C to collect the high speed supernatant (S) & microvesicle pellet (M). Samples were solubilized at 95 °C in SDS-sample buffer, resolved by SDS-PAGE & transferred to nitrocellulose. Portions of the blot were probed with 6D11 anti-SLO, anti-IFN gamma R1, 4B4F12 anti-CD14, 76B357.1 anti-TLR4, O91B8 anti-MyD88, 1H4B01 anti-Trif, EPR4477 anti-Alkaline Phosphatase, MANLAC-4A7 anti-Lamin A/C, & AC-15 anti-beta -Actin antibodies followed by relevant secondary antibodies & ECL. Full-length blots are presented in Supplementary Figure S1. (B) BMDM were challenged with 500 HU/mL SLO WT or SLO N402C, SLO ML at equivalent mass to SLO WT, or SLO N402E at equivalent mass to SLO N402C for 10 min. Cell pellets (C), high speed supernatants (S) & microvesicles (M) were isolated as in (A) & probed with 6D11 anti-Streptolysin O, 76B357.1 anti-TLR4, O91B8 anti-MyD88, 1H4B01 anti-Trif, EPR4477 anti-Alkaline Phosphatase, MANLAC-4A7 anti-Lamin A/C, & AC-15 anti-beta -Actin antibodies. Full-length blots are presented in Supplementary Figure S2. The blots are representative of at least 3 independent experiments. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-24955-2), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Pericryptal Myofibroblasts are Responsible for Increased TLR4 Expression in a Subset of CRCs. A) CRCs were separated into two groups representing low- & high- stromal expression of TLR4 by IHC staining. In normal tissue, stromal TLR4 expression is mainly due to macrophages (Green: TLR4, Red: CD68, Merge: TLR4 + CD68 + DAPI (blue)). Conversely, in CRCs increased vimentin & decreased CD68 staining in the pericryptal space confirm that this signal was due to pericryptal myofibroblasts & not related to tumor-associated macrophages. B) Double-stained immunofluorescence for TLR4 (green) & vimentin (red) in normal (I), adenoma (II), & colon adenocarcinoma (III) (10×). In the stromal compartment of CRCs, immunofluorescent staining for TLR4 localized to the pericryptal myofibroblasts in a subset of samples. C) IHC staining of colon adenocarcinoma for TLR4, vimentin, & alpha -SMA (40×). Staining co-localizes to the pericryptal space, confirming the signal arises from pericryptal myofibroblasts. D, E, & F) An increase in IHC staining for alpha -SMA & vimentin was noted in CRCs when compared to normal or low grade dysplasia. A decrease in staining for CD68 positive macrophages was observed with higher degrees of dysplasia. Image collected & cropped by CiteAb from the following publication (//jeccr.biomedcentral.com/articles/10.1186/1756-9966-33-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - (a1) Q-PCR assay of TLR4 mRNA expression levels after transfecting JJN3 cell line with TLR4 RNAi oligonucleotides or a non-targeting pool (siCtrl) for 48 h. (a2) Representative immunoblotting analyses of protein samples probed with an antibody against TLR4 after TLR4 RNAi for 48 h. (b1) % cell viability & (b2) % proliferation of JJN3 cells after TLR4 RNAi for 48 h. (c,d1) Q-PCR expression analyses of TLR4 mRNA expression levels after transfecting H929 (c) & U266 (d1) cell lines with the pCMV6-TLR4 construct or a pCMV6 empty vector for 48 h. (d2) Immunobloting analyses of U266 cells transfected with pCMV6-TLR4 or pCMV6 vector for 48 h; protein samples were probed with an antibody against TLR4. (e1) % cell viability & (e2) % proliferation of H929 & U266 cells after transfection with the pCMV6-TLR4 construct or with the empty pCMV6 vector for 48 h. beta -ACTIN probing & beta -ACTIN mRNA expression were used as reference for total protein & mRNA input, respectively. Cells transfected with the pCMV6-TLR4 construct are labelled as TLR4 OE (TLR4 overexpression), whereas cells transfected with the pCMV6 empty vector are labelled as Con (control). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30824741), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Dasatinib affects TLR4 signaling to regulate LPS-induced proinflammatory cytokine levels in BV2 microglial cells. a BV2 microglial cells were pretreated with TAK-242 (a TLR4 receptor inhibitor, 500 nM) or vehicle (1% DMSO) for 30 min, treated with dasatinib (250 nM) or vehicle (1% DMSO) for 30 min, & treated with LPS (1 μg/ml) or PBS for 5 h. The mRNA levels of COX-2 & IL-6 were analyzed by RT-PCR. b, c Quantification of the data in a (COX-2: con, n = 10; LPS, n = 10; dasatinib+LPS, n = 10; TAK-242+LPS, n = 10; TAK-242+dasatinib+LPS, n = 10; & IL-6: con, n = 14; LPS, n = 14; dasatinib+LPS, n = 14; TAK-242+LPS, n = 14; TAK-242+ dasatinib+LPS, n = 14). d BV2 microglial cells were treated with dasatinib (250 nM) or vehicle (1% DMSO) for 30 min followed by LPS (1 μg/ml) or PBS for 5.5 h, & cell surface biotinylation was conducted. e, f Quantification of the data in d (Surface TLR4: con, n = 11; LPS, n = 11; dasatinib+LPS, n = 11; Total TLR4: con, n = 16; LPS, n = 16; dasatinib+LPS, n = 16). One-way ANOVA with Tukey’s post hoc test was used to analyze significant differences. **p < 0.01, ***p < 0.001 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31655606), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - TLR4 expression in pVW-MSCs cultured with or without LPS (10 μg/ml) for 1 & 4 h & after additional 24 h of recovery after LPS removal (4hR). a: flow cytometry analysis were performed in not fixed & not permeabilized cells for TLR4 surface expression determination (TLR4: Surface) & in fixed & permeabilized cells to measure the overall TLR4 amount (TLR4: Total). Red histograms: stained cells; blue histograms: control cells. b: representative Western Blot of TLR4 & housekeeping beta -tubulin & relative quantification were presented. c: representative images of TLR4 immunostaining of pVW-MSCs cultured with or without LPS (10 μg/ml) for 4 h. pVW-MSCs nuclei were stained with Hoechst 33258 (blue). Scale bar = 10 μm. Data shown represent the mean ± SD of three biological replicates, each experiment is repeated three times. Data were analysed using one-way ANOVA followed by the Tukey’s post hoc comparison test. Different letters above the bars indicate significant differences (p < 0.05). (AU = Arbitrary Units) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31029157), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Flow Cytometry: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - TLR4 expression in pVW-MSCs cultured with or without LPS (10 μg/ml) for 1 & 4 h & after additional 24 h of recovery after LPS removal (4hR). a: flow cytometry analysis were performed in not fixed & not permeabilized cells for TLR4 surface expression determination (TLR4: Surface) & in fixed & permeabilized cells to measure the overall TLR4 amount (TLR4: Total). Red histograms: stained cells; blue histograms: control cells. b: representative Western Blot of TLR4 & housekeeping beta -tubulin & relative quantification were presented. c: representative images of TLR4 immunostaining of pVW-MSCs cultured with or without LPS (10 μg/ml) for 4 h. pVW-MSCs nuclei were stained with Hoechst 33258 (blue). Scale bar = 10 μm. Data shown represent the mean ± SD of three biological replicates, each experiment is repeated three times. Data were analysed using one-way ANOVA followed by the Tukey’s post hoc comparison test. Different letters above the bars indicate significant differences (p < 0.05). (AU = Arbitrary Units) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31029157), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Toll‐like receptor 4 (TLR4)‐shRNA lentivirus reduced myocardial inflammation & improved heart function after myocardial infarction (MI). The rats received intra‐myocardial injection of normal saline (NS), control‐shRNA lentivirus or TLR4‐shRNA lentivirus (1 × 109 TU/ml, 100 μl/heart) just after left anterior descending coronary artery (LAD) ligation or sham operation. All examinations were performed after 4 weeks of MI. (A) Expression of green fluorescent protein (GFP; green), the marker gene carried by TLR4‐shRNA lentivirus, in the myocardium. The nuclei were counter‐stained with Hoechst 33258 (blue). (B) Representative Western blot images & quantification of TLR4 proteins in sham & chronic heart failure (CHF) myocardium. (C) tumour necrosis factor (TNF)‐ alpha & interleukin (IL)‐6 protein contents in infarct & remote myocardium. (D) Representative images of Masson's trichrome staining (upper panel) & quantification (lower panel) of post‐infarct failing hearts, showing that TLR4‐shRNA lentivirus reduced cardiac fibrosis. Cross‐sections were cut at the midhorizontal plane of the fixed paraffin‐embedded heart, & stained with Masson's trichrome reagents. (E) Infarct size of post‐infarct failing hearts. (F) Fractional shortening (%) of the left ventricle (data are means ± SD, n = 4/group, aP < 0.05, AP < 0.01 versus respective sham‐NS; BP < 0.01 versus respective CHF‐NS). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26290459), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - ALWPs significantly reduced LPS-induced cell-surface levels of TLR4. (A) BV2 microglial cells were pretreated with TAK-242 (a TLR4 inhibitor, 500 nM) for 30 min, followed by treatment with ALWPs (500 μg/ml) or PBS for 30 min & finally LPS (1 μg/ml) or PBS for 5 h. Total RNA was isolated, & IL-1 beta mRNA levels were measured by RT-PCR. (B) Quantification of the data from (A) (con, n = 20; LPS, n = 20; ALWPs + LPS, n = 20; TAK-242 + LPS, n = 20; TAK-242 + ALWPs + LPS, n = 20). (C) BV2 microglial cells were pretreated with ALWPs (500 μg/ml) or PBS for 30 min, followed by treatment with LPS (1 μg/ml) or PBS for 5.5 h. Cell-surface biotinylation was then conducted with a TLR4 antibody recognizing the N-terminal region of TLR4. (D) Quantification of the data from (C) (Surface TLR4: con, n = 10; LPS, n = 10; ALWPs + LPS, n = 10; Total TLR4: con, n = 4; LPS, n = 4; ALWPs + LPS, n = 4). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30319390), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ma, Pm, ChHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ma, Pm, ChHa

Applications:

, , , , , , , , , , , , , , ,

Applications: , , , , , , , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-5031A

Alt Catalog# IMG-5031A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-27149AF350
Alexa Fluor 405	NBP2-27149AF405
Alexa Fluor 488	NBP2-27149AF488
Alexa Fluor 532	NBP2-27149AF532
Alexa Fluor 594	NBP2-27149AF594
Alexa Fluor 647	NBP2-27149AF647
Alexa Fluor 700	NBP2-27149AF700
Alexa Fluor 750	NBP2-27149AF750
Allophycocyanin	NBP2-27149APC
Allophycocyanin/Cy7	NB100-56566APCCY7
Biotin	NBP2-27149B
DyLight 350	NBP2-27149UV
DyLight 405	NBP2-27149V
DyLight 488	NBP2-27149G
DyLight 550	NBP2-27149R
DyLight 594	NB100-56566DL594
DyLight 650	NBP2-27149C
DyLight 680	NBP2-27149FR
DyLight 755	NBP2-27149IR
FITC	NBP2-27149F
HRP	NBP2-27149H
Janelia Fluor 549	NB100-56566JF549
Janelia Fluor 646	NB100-56566JF646
PE	NBP2-27149PE
PE/Atto594	NB100-56566PEATT594
PE/Cy5.5	NB100-56566PECY55
PE/Cy7	NB100-56566PECY7
PerCP	NBP2-27149PCP
mFluor Violet 450 SE	NBP2-27149MFV450
mFluor Violet 500 SE	NBP2-27149MFV500
mFluor Violet 610 SE	NBP2-27149MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Azide and BSA Free	NBP2-27149
BSA Free	NB100-56566SS
Azide and BSA Free	NBP2-27149SS

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NB300-144

Laminin Antibody - BSA Free

(8 Reviews)

(140 Publications)

0.1 ml

NB300-144

MxIF analysis of mouse submandibular salivary gland morphogenesis. MxIF of a developmental TMA including embryonic stages (E14, E16, E18) and postnatal stages (P5 and P20) was performed using sequential application of directly conjugated antibodies to detect multiple markers of tissue structures and cell types on the same tissue sections. Tissue compartments. The epithelium, mesenchyme, neurons, and basement membranes was detected using antibodies directed towards E-cadherin (ECAD, red), platelet-derived growth factor (PDGFR, green), beta III tubulin (bIII, magenta), and Laminin Antibody [NB300-144] (LMN, cyan), respectively. Image collected and cropped by CiteAb from the following publication (//bio.biologists.org/cgi/doi/10.1242/bio.20134309), licensed under a CC-BY license.

Staining of mouse section of cortex stained with Laminin Antibody [NB300-144] (red). Blue is DAPI staining of DNA. This antibody reveals strong staining in the basement membranes of blood vessels.

Staining of rat spinal cord and dorsal root paraformaldehyde/paraffin-embedded tissue using Laminin Antibody [NB300-144]. Pepsin antigen retrieval was performed on this tissue sample.

Analysis of rat heart cells lysates (lane 1) and 0.2 ug of purified laminin protein from mouse EHS sarcoma (lane 2) Laminin Antibody [NB300-144] recognizes 3 laminin isotypes: alpha 1 (440 kDa), beta 1 (220 kDa) and gamma 1 (220 kDa). Also recognized is a laminin binding protein at 120 kDa in both rat heart lysates and purified laminin protein. Since this protein always coexpresses with laminin this crossreactivity is irrelevant. Theoretical molecular weight of LAMA1 is 337 kDa.

IF Confocal analysis of HeLa cells using Laminin Antibody [NB300-144] (1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green, A). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red, B). DAPI was used to stain the cell nuclei (blue, C).

Western blot analysis of the effects of IP6, Ins, IP6 + Ins and normal saline on the levels of collagen IV, Laminin and Fibronectin, with Laminin Antibody [NB300-144]. IP6 or Ins treatment decreased the protein expression of LN and FN, and the combined IP6 + Ins treatment resulted in significantly greater effects compared with treatment with either compound alone. The Western blot membranes were stripped and reprobed for beta-actin as an internal control to confirm equal loading. (A) representative blots from one of three separate experiments; (B) relative band intensities based on densitometry. The results are expressed as the mean +/- standard deviation from three independent experiments. * p < 0.05 compared to the IP6 + Ins group; #p < 0.05 compared to the normal saline group. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2072-6643/8/5/286), licensed under a CC-BY license.

Immunofluorescence staining for extracellular matrix proteins in day 1 and day 7 HWJSC spheroids. HWJSC spheroids were cultured for either 1 day or 7 days (with 6 days of osteogenic differentiation), fixed, and stained for extracellular matrix protein: laminin (C) and counter-staining with Hoechst. Scale bars represent 100 um. Image collected and cropped by CiteAb from the following publication (journals.plos.org/plosone/article?id=10.1371/journal.pone.0184155), licensed under a CC-BY license. Using the Alexa Fluor 488 format of this antibody.

Laminin staining on an E12.5 mouse Right Ventricle. IHC-Fr image submitted by a verified customer review.

A surface stain was performed on HeLa cells with the Alexa Fluor 647 conjugate of Laminin Antibody [NB300-144AF647] (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 2.5 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Analysis of Laminin-1 expression in mouse EHS tumor crude extracts (left) and Laminin-2 expression in rat heart crude extracts (right), using Laminin Antibody [NB300-144]. The Laminin polyclonal antibody was used at 1 ug/mL. Theoretical Molecular Weight of NB300-144 is 337 kDa.

VSOP observed in perivascular-restricted spinal cord lesions with intact BBB. Immunostaining for laminin (brown) using Laminin Antibody [NB300-144] shows vascular endothelium and glia limitans of a perivascular lesion, along with infiltrating cells and VSOP (blue). Image collected and cropped by CiteAb from the following publication (//asn.sagepub.com/lookup/doi/10.1042/AN20120081), licensed under a CC-BY license.

SOX17 regulates adult muscle regeneration after injury in Pax7CreERT2/+;Sox17fl/fl mutant mice. Representative images of cryosections from regenerating adult TA muscles d28 after injury, showing immunofluorescence for PAX7+ (quiescent, arrows) cells. Scale bar, 25 Aum. Image collected and cropped by Citeab from the following publication (SOXF factors regulate murine satellite cell self-renewal and function through inhibition ofI2-catenin activity. Elife (2018)) licensed under a CC-BY license.

WNT11 regulates MMPs activities.(A–C) (Top panels): Serum-free medium was conditioned for 24 hrs by the indicated cells, concentrated 20-fold & assayed by gelatin zymography. Gelatinolytic activity is indicated by clear zones against a dark background of stained substrate. (Bottom): Whole cell extracts were immunoblotted with indicated antibodies. (A) Overexpression of Wnt11 in EMSC or BT473 cells enhances activity of MMP-9 & MMP-2. (B) Impaired activity of MMP-9 & MMP-2 in MDA-MB-231 cells (left) or EMSCs (right) stably expressing Wnt11 shRNAs & treated with DMOG. (C) WNT11 is required for MMP-9 & MMP-2 activity in MDA-MB-231 cells (left) or EMSCs (right) under normoxic & hypoxic culture conditions. (D) WNT11 regulates MMP2 protein in media. (Top): conditioned media from indicated cells & treatments. (Bottom): whole cell lysates were immunoblotted with indicated antibodies. (E) Recombinant WNT11 induces both MMP-2 protein & MMP-2 activity in media. (Top panels): Gelatin zymography & immunoblot of serum-free medium conditioned for the indicated times after recombinant WNT11 (r-WNT11) treatment. (Bottom): Whole cell lysates were immunoblotted with indicated antibodies. (F) MMP-2 inhibitor attenuated induced migration by WNT11. MDA-MB-231 cells infected with lentiviruses for stable expression of Wnt11 or GFP (n = 4) were incubated with either vehicle or 1 μM of ARP100. Media in the lower compartment had same concentration of DMSO or inhibitor. Values are mean ± s.e.m. *p < 0.05, **p < 0.01. For panels (A–D), HIF-1 alpha & HIF-2 alpha were shown as a marker of hypoxia, WNT11 normalized to alpha -Tubulin was shown. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Hypoxia induces expression of WNT11 through VHL.(A,B) Higher basal levels of WNT11 protein in Vhl-deleted cells (lenti-Cre infected Vhlf/f). EMSCs isolated from Vhlf/f mouse were infected with lentivirus carrying either GFP gene (for control) or Cre recombinase (for knockout). Non-infected cells were also used as a control. Immunoblot analysis of control or Vhl KO EMSCs treated with 0.1 mM DMOG (A), & EMSCs exposed to air (21% O2) or hypoxia (1% O2) for 24 hrs (B). Laminin, alpha -tubulin, & lamin A/C were used as loading controls, WNT11 normalized to alpha -Tubulin was shown. (C,D) Inactivation of the Vhl gene results in increased Wnt11 mRNA. Wnt11 & Vegf mRNA levels in liver (C) or duodenum (D) were measured by qPCR in Liver-VhlcKO or duodenum-VhlcKO & control mice (n = 5 per group). Values normalized to Tbp mRNA are expressed relative to tissues from control mice. For panels (C,D), values are mean ± s.e.m. *p < 0.05, **p < 0.01. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

WNT11 is induced by hypoxia or hypoxic mimetics in different cell types.(A) Increased Wnt11 mRNA in EMSC adipocytes (Day 12) after hypoxia-mimetic treatments. EMSC adipocytes were treated with CoCl2 (0.1 mM), DFO (0.1 mM) or DMOG (0.1 mM) for 24 hrs. Values were normalized to Tbp mRNA & are expressed relative to control (n = 3). (B,C) Increased Wnt11 mRNA by hypoxia in EMSC preadipocytes & adipocytes (Day 0–12 after differentiation) (B), & C2C12 myoblast & myocyte (Day 0 & 8 after differentiation) (C). Wnt11 mRNA was assessed by quantitative PCR in cells exposed to air (21% O2) or hypoxia (1% O2) for 24 hrs. (n = 4). Values were normalized to Tbp mRNA & are expressed relative to 21% O2 samples (left panel). (D) Immunoblot analyses of HeLa cells under normal air or hypoxia for 24 hrs. (E,F) Induction of Wnt11 by increasing concentrations of DMOG in MDA-MB-231 cells (E) & 4T1 cells (F). (G) EMSCs treated with 0.1 mM DMOG for the indicated times. Wnt11 & Vegf mRNA expression was measured by qPCR & normalized to Tbp mRNA (n = 4). (H) WNT11 protein levels after DMOG treatment normalized to alpha -Tubulin (upper panel; n = 4). Representative immunoblots of EMSCs treated with 0.1 mM DMOG for the indicated times (Lower panel). (I) Protein expression in MDA-MB-231 cells treated with 0.1 mM DMOG. (J) Induction of Wnt11 promoter activity by hypoxia or hypoxia mimetics. pGL3-Wnt11 promoter plasmid was transfected into C2C12 cells. Cells were incubated with DMOG (left panel, n = 4) or under 21% O2 or 1% O2 (right panel, n = 8) for 24 hrs. For panels (A–C,G,H,J), values are mean ± s.e.m. *p < 0.05, **p < 0.01. For panels of immunoblotting, laminin, alpha -tubulin, & ERK were used as loading controls, WNT11 normalized to alpha -Tubulin was shown. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Laminin Antibody [NB300-144] - hS/PCs sequentially secrete basement membrane proteins during microstructure reorganization & function. Encapsulated hS/PCs in HA-PEGDA hydrogels initially produce laminin (red) (A–C) & collagen IV (green) (D–F) even at the single-cell state (A,D). Perlecan (green) (G–I) secretion follows to stabilize the laminin & collagen IV networks. Scalebar = 20 μm in all confocal micrographs. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31750298), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Laminin Antibody [NB300-144] - Western blot analysis of the effects of IP6, Ins, IP6 + Ins & normal saline on the levels of collagen IV, Lamininand Fibronectin. IP6 or Ins treatment decreased the protein expression of collagen IV, LN & FN, & the combined IP6 + Ins treatment resulted in significantly greater effects compared with treatment with either compound alone. The samples were probed with antibodies against p-collagen IV, p-LN, & p-FN. The Western blot membranes were stripped & reprobed for beta -actin as an internal control to confirm equal loading. (A) representative blots from one of three separate experiments; (B) relative band intensities based on densitometry. The results are expressed as the mean ± standard deviation from three independent experiments. * p < 0.05 compared to the IP6 + Ins group; #p < 0.05 compared to the normal saline group. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/2072-6643/8/5/286), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, ChHa, In, Ma, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, ChHa, In, Ma, Rb, Sh

Applications:

, , , ,

Applications: , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB300-144AF350
Alexa Fluor 405	NB300-144AF405
Alexa Fluor 488	NB300-144AF488
Alexa Fluor 532	NB300-144AF532
Alexa Fluor 594	NB300-144AF594
Alexa Fluor 647	NB300-144AF647
Alexa Fluor 700	NB300-144AF700
Alexa Fluor 750	NB300-144AF750
Allophycocyanin	NB300-144APC
Allophycocyanin/Cy7	NB300-144APCCY7
Biotin	NB300-144B
CoraFluor 1	NB300-144CL1
DyLight 350	NB300-144UV
DyLight 405	NB300-144V
DyLight 488	NB300-144G
DyLight 550	NB300-144R
DyLight 594	NB300-144DL594
DyLight 650	NB300-144C
DyLight 680	NB300-144FR
DyLight 755	NB300-144IR
FITC	NB300-144F
HRP	NB300-144H
Janelia Fluor 525	NB300-144JF525
Janelia Fluor 549	NB300-144JF549
Janelia Fluor 646	NB300-144JF646
Janelia Fluor 669	NB300-144JF669
PE	NB300-144PE
PE/Atto594	NB300-144PEATT594
PE/Cy5.5	NB300-144PECY55
PE/Cy7	NB300-144PECY7
PerCP	NB300-144PCP
mFluor Violet 450 SE	NB300-144MFV450
mFluor Violet 500 SE	NB300-144MFV500
mFluor Violet 610 SE	NB300-144MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB300-144SS

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NB600-503

beta-Actin Antibody - BSA Free

(4 Reviews)

(135 Publications)

0.05 ml

NB600-503

Simple Western: beta-Actin Antibody [NB600-503] - Simple Western lane view shows a specific band for Beta Actin in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Flow Cytometry: beta-Actin Antibody [NB600-503] - An intracellular stain was performed on HeLa cells with beta-actin Antibody NB600-503AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: beta-Actin Antibody [NB600-503] - Western blot analysis of Jurkat, COS, and CHO cell lysate using beta actin antibody at 1:100.

Immunohistochemistry-Paraffin: beta-Actin Antibody [NB600-503] - Analysis of Beta Actin in mouse epidermis using DAB with hematoxylin counterstain.

Immunocytochemistry/Immunofluorescence: beta-Actin Antibody [NB600-503] - IF Confocal analysis of HeLa cells using Beta Actin antibody (NB600-503, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). DAPI was used to stain the cell nuclei (blue).

Flow Cytometry: beta-Actin Antibody [NB600-503] - Analysis of HeLa cells using mouse Monoclonal beta-Actin antibody (Orange) and Isotype control Antibody (Blue).

Simple Western: beta-Actin Antibody [NB600-503] - Early start GM1 administration did not affect alpha-synuclein expression or transport to the striatum. When assessed 1 week following AAV-A53T alpha-synuclein injection, levels of striatal alpha-synuclein were no different in saline (N=6) vs. GM1-treated animals (N=6), suggesting no influence of GM1 on A53T alpha -synuclein transduction or transport to the striatum. Representative Wes Simple Western blots are shown after cropping (full length images of blots are presented as Supplementary Fig. 2. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-019-42847-x), licensed under a CC-BY license.

Western Blot: beta-Actin Antibody [NB600-503] - Regulation of inflammatory signaling in hSMC.ND (open bars) & T2D (solid bars) cells extracted after 48 hr treatment with LPS, Pioglitazone (Pio), palmitate or oleate. Results expressed relative to untreated control for each individual set of cells, Ave + SEM. (A) IkBa protein, n = 7–12 & 6–15 for ND & T2D, respectively. (B) Total & phospho-p38, n = 6–14 & 5–8. (C) Total & phospho-p44/42, n = 7–13 & 7–11. (D) Total & phospho-JNK, n = 9–12 & 3–8. * p<0.05 T2D vs ND. †p<0.05 T2D response vs ND response Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27453994), licensed under a CC0-1.0 license. Not internally tested by Novus Biologicals.

Western Blot: beta-Actin Antibody [NB600-503] - Impact of T2D on inflammatory signaling in hSMC.Proteins extracted from ND (open bars) & T2D (solid bars) hSMC under control conditions. (A) Representative western blots. (B) Quantification of protein expression normalized to loading control ( beta -actin). Ave + SEM, n = 13–21 for ND, 10–16 for T2D. (C) Protein phosphorylation, expressed as ratio of phosphorylated to total protein. Ave + SEM, n = 13–17 for ND, 9–15 for T2D. *p<0.05 vs ND Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27453994), licensed under a CC0-1.0 license. Not internally tested by Novus Biologicals.

Western Blot: beta-Actin Antibody [NB600-503] - Early start GM1 administration did not affect alpha -synuclein expression or transport to the striatum. (A,B) When assessed 1 week following AAV-A53T alpha -synuclein injection, levels of striatal alpha -synuclein were no different in saline (N = 6) vs. GM1-treated animals (N = 6), suggesting no influence of GM1 on A53T alpha -synuclein transduction or transport to the striatum. Representative Wes Western blots are shown after cropping (full length images of blots are presented as Supplementary Fig. 2. (C) Double label immunofluorescence 1 week after AAV-A53T alpha -synuclein injection showed no differences between saline & GM1-treated animals in alpha -synuclein accumulation (green) in TH+ neurons (red) in the SNc. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31182727), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: beta-Actin Antibody [NB600-503] - Anti-hnRNP A1 antibodies alter protein levels as measured by Western blot. SK-N-SH cells were cultured & treated with anti-hnRNP A1 antibodies or control IgG. Following a 48 hour incubation, cells were lysed & protein lysate was run on 10% Tris-glycine gels for Western blot analysis & probed for Beta-actin (A) or GAPDH (B)) (control), hnRNP A1, SPG4, SPG7, & SPG20. Results revealed that there was a marked reduction of SPG 4 & SPG 7 protein levels in anti-hnRNP A1 antibody compared to control isotype IgG treated cells. There was a variable response to hnRNP A1 protein & a modest reduction of SPG 20. Parentheses show relative percent reduction of signal comparing anti-hnRNP A1 antibody to control isotype IgG treatment of cells. (L=lysate, IgG=control isotype IgG, A1=anti-hnRNP A1-M9 antibody treatment of cells). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27375925), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Av, Bv, ChHa, Fi, Pm, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Av, Bv, ChHa, Fi, Pm, Rb

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 405	NB600-503AF405
Alexa Fluor 488	NB600-503AF488
Alexa Fluor 647	NB600-503AF647
Alexa Fluor 700	NB600-503AF700
Allophycocyanin	NB600-503APC-0.05ml
Biotin	NB600-503B
DyLight 350	NB600-503UV-0.05ml
DyLight 405	NB600-503V
DyLight 488	NB600-503G
DyLight 550	NB600-503R
DyLight 594	NB600-503DL594
DyLight 650	NB600-503C
DyLight 680	NB600-503FR-0.05ml
DyLight 755	NB600-503IR-0.05ml
FITC	NB600-503F
HRP	NB600-503H
Janelia Fluor 549	NB600-503JF549
Janelia Fluor 646	NB600-503JF646
PE	NB600-503PE-0.05ml
PerCP	NB600-503PCP-0.05ml

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-503SS

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

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Modification

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Clone

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Species Glossary

LC3B Antibody - BSA Free

GAPDH Antibody (1D4) - BSA Free

ABCA1 Antibody - BSA Free

SR-BI Antibody - BSA Free

Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free

ABCG1 Antibody - BSA Free

p62/SQSTM1 Antibody - BSA Free

TRF-2 Antibody - BSA Free

alpha Tubulin Antibody (DM1A) - BSA Free

TLR4 Antibody (76B357.1) - BSA Free

Laminin Antibody - BSA Free

beta-Actin Antibody - BSA Free