Neuroscience reagents: Novus Biologicals

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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NB600-1384

LC3B Antibody - BSA Free

(19 Reviews)

(459 Publications)

0.1 ml

NB600-1384

FTC-133/TPC-1 cells were transfected with HMGB1 shRNA and control shRNA and then pre-treated for 1 h with pepstatin A (PA, 10 uM) and E64D (10 uM) as indicated. Cells were subsequently treated for 3 h with HBSS in continuous presence or absence PA/E64D inhibitors. LC3-I/II levels were assayed by Western blot. Image collected and cropped by CiteAb from the following publication (//jeccr.biomedcentral.com/articles/10.1186/s13046-019-1328-3) licensed under a CC-BY license.

LC3B immunohistochemical staining in CC tissue(A) CC with low LC3B dot like staining. (B) CC with high LC3B dot like staining. (magnification 20X). Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17554) licensed under a CC-BY license.

Analysis of LC3B in treated U87-MG (human glioblastoma astrocytoma) lysates using anti- [Catalog # NB600-1384].

Depletion of RARalpha prevents ATRA-induced autophagic flux in SKBR3 cells. LC3B western blotting and quantification of the autophagic activity in control and RARalpha-knockdown SKBR3 cells upon treatment with 1 uM ATRA for 2 days in the presence or absence of BafA for 2 h at 200 nM. LC3B-II expression was normalized to GAPDH and to vehicle-treated control cells. Standard deviations from at least five independent experiments are shown. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2015236), licensed under a CC-BY license.

Human cervical carcinoma (HeLa) and Mouse Neuroblast cells (Neuro2a) were treated with (+) and without (-) 50 uM Chloroquine overnight. Whole cell protein lysates were prepared in 1x Laemmli sample buffer and approximately 10 ug of each lysate (NBP2-49689 and NBP2-49688) was separated on a 4- 20% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% nonfat milk in TBST. The membrane was probed with 2 ug/mL anti-LC3 (NB600-1384) and 1 ug/mL anti-alpha tubulin (NB100-690) as a loading control, and detected with the appropriate secondary antibodies using NovaLume Pico Chemiluminescence Substrate (NBP2-61915).

<b>Biological Strategies Validation. </b>Fibroblasts were exposed to 1 mM LLOMe and when indicated pretreated with the autophagy inhibitors 3-methyladenine (5 mM, 1 h) or bafilomycin A1 (25 nM, 30 min). Immunostaining with corresponding colocalization analysis of a) LC3 (green) and galectin-3 (red) and b) LC3 (green) and LAMP2 (red). Merged images show colocalization in yellow, nuclei are stained with DAPI (blue). LLOME = l-leucyl-l-leucine methyl ester, a lysosomal membrane damage compound. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32409651/) licensed under a CC-BY license.

Confocal analysis of HeLa cells using Rabbit anti-LC3B antibody (Catalog # NB600-1384, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).

Immunocytochemical/Immunofluorescent staining of treated U373-MG cells using the HRP conjugate of anti- (Catalog # NB600-1384). The nuclei were stained with DAPI.

Staining of treated U373-MG (human glioblastoma) cells using anti- [Catalog # NB600-1384].

Analysis of U87MG glioma xenografts using anti- [Catalog # NB600-1384]. Image from verified customer review.

LC3B staining in treated U87-MG cultured & subcutaneous tumors.

LC3B staining in glioblastoma multiform tissue.

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

LC3A and LC3B siRNAs specifically block the expression of the LC3A and LC3B proteins, respectively, in A549 cell line (E1,2,3). In E4 the reactivity of and of the LC3B (5F10 antibody) is shown, following silencing of the LC3A or of the LC3B genes, in the A549 cell line. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0137675), licensed under a CC-BY license.

NOD1-dependent induction of autophagy contributes to IRGM and LC3 recruitment to Mtb-containing vesicles. The subcellular localization of autophagy proteins was observed in untreated (A, C) and Tri-DAP treated cells (B, D) using anti-IRGM and anti-LC3 antibodies, which were detected using a secondary antibody coupled to 5 nm gold particles (indicated with arrowheads; TEM X 62,000). Gold particles colocalizing with bacteria were manually counted in 10 macrophages of each condition (E, F) and the differences between the treatments are indicated: ***p 0.05 versus TBI + vehicle group. beta -actin was used as a loading control. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28429775), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Differences in neuronal autophagy & dendrite varicosity following HIV-1 Tat protein & morphine treatment. (A) Representative images of neurons transfected with a fluorescent reporter plasmid to monitor autophagic flux at 8 h following the indicated treatments. GFP (green) & GFP + mRFP (yellow) fluorescence are observed prior to the fusion of autophagosomes with lysosomes whereas only mRFP (red) fluorescence is present in post-fusion autolysosomes. DIC, differential interference contrast microscopy image. DAPI (blue) staining indicates cell nuclei. (B) Quantification of autolysosomes (red puncta) from (A). F(3,13) = 8.756, p = 0.0019; ∗p 0.05 versus TBI + DMSO group. beta -actin was used as a loading control. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28429775), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

8-Cl-Ado-induces autophagic cell killing. (A) Western blot analysis of beclin1 & ATG7 levels in MCF-7 cells transfected with either a pool of control siRNA (siCONT), siRNA targeting the expression of the beclin1 gene (siBECN1), or targeting the expression of the ATG7 gene (siATG7). Immunoblot analysis of LC3B lipidation & PARP cleavage were assessed as markers of autophagosome formation & apoptosis, respectively. GAPDH was used as loading control. Flow cytometric analysis of cells transfected with siCONT, solid bars, siBECN1, hatched bars, or siATG7, checkered bars, treated with 10 μM 8-Cl-Ado & stained with (B) annexin V & PI, as well as (C) acridine orange. Effect of autophagy on 8-Cl-Ado-inhibiton of clonogenic survival. Cells transfected with (D) siCONT, ○, or siBECN1, ●, & with (E) siCONT, ○, or siATG7, ●, were treated with the indicated doses of 8-Cl-Ado for 3 days, washed with PBS, & cultured in fresh medium for 10 days. Colonies of >50 cells were counted under a dissecting microscope. Image collected & cropped by CiteAb from the following publication (//jhoonline.biomedcentral.com/articles/10.1186/1756-8722-7-23), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LC3B Antibody - BSA Free [NB600-1384] - Assesment of functional basal autophagy in vitro(A) Immunoblotting of LC3B & p62 upon pharmacological autophagy inhibition. A dose-dependent increase of LC3B & p62 was observed with increasing concentrations of chloroquine. (B) Immunofluorescence of LC3B & p62 upon pharmacological autophagy inhibition. I. In CC cell lines HT-29 & II. LoVo a dose-dependent increase of LC3B & p62 was observed with increasing concentrations of chloroquine. (C) Electron microscopy of CC cell lines pharmacological autophagy inhibition. I-III. In CC cell line HT-29 a dose-dependent increase of cytoplasmic vesicles was observed with increasing concentrations of CQ. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28903368), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - Phosphorylation of AKT & mTOR decreases after treatment with gemcitabine (GCB)A LM7, CCH-OS-D, & K7M3 osteosarcoma cells were treated with 1μM GCB & cell lysates were analyzed by immunoblotting with specific antibodies for AKT, mTOR, p-AKT, & p-mTOR. beta -actin was used as a loading control. Densitometry was performed using ImageJ. B. & C. Following treatment with GCB, lysates were analyzed using the Human Phospho Kinase Antibody Array. The phosphorylation levels of AKT, mTOR, PRAS40, p70S6K, JNK, AMPK, & ERK were evaluated. Densitometry was performed using Image J. Values were graphed as relative fold change. Means ± standard deviation of three independent experiments are shown. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.20308), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - Autophagy decreases during striatal development. (A) Representative Western blot images for LC3B, p62, DARPP32, & actin. Quantification of (B) LC3B-ii relative to actin, (C) LC3B-ii relative to LC3B-i, (D) p62 relative to actin normalized to P120 values. Data analyzed with one-way ANOVA; (B) Age: F(4,21) = 6.526, p = 0.0014; (C) Age: F(4,22) = 3.797, p = 0.0171; (D) Age: F(4,15) = 3.762, p = 0.0260. *p 0.01); (b) FTC-133/TPC-1 cells transfected with HMGB1 shRNA & control shRNA were starved by HBSS for 3 h & then treated with rapamycin (1 μM) for 12 h. Flow cytometry was performed for measuring the ROS level by a DCFH-DA probe in indicated cells. Rap, rapamycin. (n = 3, *P 0.01); (c) FTC-133/TPC-1 cells were transfected with HMGB1 shRNA & control shRNA & then starved by HBSS for 3 h. ATP levels were detected by ATP Assay Kit (n = 3, *P > 0.01); (d) FTC-133/TPC-1 cells were pretreated with NAC (2 mM) & then starved by HBSS for 3 h. LC3-I/II, NIS, p-AMPK, AMPK, p-mTOR, mTOR, p-p70S6K & p70S6K levels were assayed by Western blot; (e) FTC-133/TPC-1 cells transfected with HMGB1 shRNA & control shRNA were starved by HBSS for 3 h & then treated with rapamycin (1 μM) for 12 h. LC3-I/II, NIS, p-mTOR & mTOR levels were assayed by Western blot. Rap, rapamycin Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31331356), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - Serum starvation induces autophagy & apoptosis in endothelial cells.A Evaluation by Hoechst 33342 & propidium iodide (HO/PI) staining of apoptosis & necrosis in HUVECs exposed to normal medium (N) or serum-starved (SS) for 4 h. P values obtained by unpaired t test. n = 10 for each condition. Scale bar = 25 μm. B Quantification of caspase-3 activity in HUVECs exposed to N or SS HUVECs for 1, 2, 3, & 4 h. P values obtained by one-way ANOVA. n = 4 for each condition. C Immunoblot & densitometric analysis of PARP, LC3, & SQSTM1/p62 in HUVECs exposed to N or SS for 1, 2, or 4 h. beta -actin (ACTB) was used as a loading control. P values obtained by unpaired t test (*P 0.01, **P > 0.01); (d) Dynamic uptaking: FTC-133/TPC-1 cells were transfected with HMGB1 shRNA & control shRNA & starved by HBSS for 3 h. Indicated cells were cultured with 175 KBq 131I for 5, l0, 20, 30, 60, 90 & 120 min. The uptake of 131I in indicated cells was detected by a gamma counter; (e) Radionuclide uptaking: FTC-133/TPC-1 cells were transfected with HMGB1 shRNA & control shRNA in the presence or absence of 3-MA (10 mM) treatment for 1 h & then starved by HBSS for 3 h. After 1-h incubating with 131I, the uptake of 131I in indicated cells was detected by a gamma counter (n = 3, *P 0.01); (f) FTC-133/TPC-1 cells transfected with HMGB1 shRNA & control shRNA were starved by HBSS for 3 h & then treated with rapamycin (1 μM) for 12 h. After 1-h incubating with 131I, the uptake of 131I in indicated cells was detected by a gamma counter. Rap, rapamycin. (n = 3, *P 0.01) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31331356), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - ROS was sufficient for inducing HMGB1 translocation & enhancing autophagy (a) FTC-133/TPC-1 cells were pretreated with the antioxidant (NAC, 2 mM) for 1 h & then starved by HBSS for 3 h. ROS production was assessed by measuring the fluorescent intensity of DCF on a fluorescent plate reader. Incremental production of ROS was expressed as a percentage of control (n = 3, *P 0.01); (b) FTC-133/TPC-1 cells transfected with HMGB1 shRNA & control shRNA were starved by HBSS for 3 h & then treated with rapamycin (1 μM) for 12 h. Flow cytometry was performed for measuring the ROS level by a DCFH-DA probe in indicated cells. Rap, rapamycin. (n = 3, *P 0.01); (c) FTC-133/TPC-1 cells were transfected with HMGB1 shRNA & control shRNA & then starved by HBSS for 3 h. ATP levels were detected by ATP Assay Kit (n = 3, *P > 0.01); (d) FTC-133/TPC-1 cells were pretreated with NAC (2 mM) & then starved by HBSS for 3 h. LC3-I/II, NIS, p-AMPK, AMPK, p-mTOR, mTOR, p-p70S6K & p70S6K levels were assayed by Western blot; (e) FTC-133/TPC-1 cells transfected with HMGB1 shRNA & control shRNA were starved by HBSS for 3 h & then treated with rapamycin (1 μM) for 12 h. LC3-I/II, NIS, p-mTOR & mTOR levels were assayed by Western blot. Rap, rapamycin Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31331356), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - Fucoxanthin activated autophagy after TBI.(A) Representative images of immunofluorescence for LC3 surrounding the injured cortex. LC3 punctate dots were observed in the cytoplasm by immunofluorescent staining of LC3 (red). Neuron cells & nuclei are labeled with NeuN (green) & DAPI (blue), respectively. Magnification: 40 x. Scale bar: 50 mm. (B) Mice brain tissues were collected 1 day after TBI in different groups, & the expression of LC3, Beclin-1 & p62 was measured by western blot. Fucoxanthin treatment significantly increased the level of LC3-II & Beclin-1 while decreasing the level of p62 after TBI. (C) 3-MA (400 nM) was injected i.c.v. 30 min before TBI. Mice were then subjected to TBI & treatment of fucoxanthin 30 min after TBI. Pretreatment with 3-MA significantly attenuated fucoxanthin-induced activation of autophagy & suppression of apoptosis & oxidative stress in the ipsilateral cortex. Data are presented as mean ± SEM, n = 6 per group; **p 0.01, **P > 0.01); (d) Dynamic uptaking: FTC-133/TPC-1 cells were transfected with HMGB1 shRNA & control shRNA & starved by HBSS for 3 h. Indicated cells were cultured with 175 KBq 131I for 5, l0, 20, 30, 60, 90 & 120 min. The uptake of 131I in indicated cells was detected by a gamma counter; (e) Radionuclide uptaking: FTC-133/TPC-1 cells were transfected with HMGB1 shRNA & control shRNA in the presence or absence of 3-MA (10 mM) treatment for 1 h & then starved by HBSS for 3 h. After 1-h incubating with 131I, the uptake of 131I in indicated cells was detected by a gamma counter (n = 3, *P 0.01); (f) FTC-133/TPC-1 cells transfected with HMGB1 shRNA & control shRNA were starved by HBSS for 3 h & then treated with rapamycin (1 μM) for 12 h. After 1-h incubating with 131I, the uptake of 131I in indicated cells was detected by a gamma counter. Rap, rapamycin. (n = 3, *P 0.01) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31331356), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - Metformin treatment increases cisplatin effects; (A) MDA-MB-231 & Ca Ski cells were either left untreated or treated with metformin or cisplatin alone or metformin in combination with cisplatin for 15 days. At the end of the treatment, cells were processed to obtain whole cell lysates. MAP1LC3B, pPRKAA2Thr172, PRKAA2 & CASP3 (cleaved) expression was determined by Western blot. ACTB was used as loading control. Densitometric analysis of the gels was performed as described under Materials & Methods; (B) MDA-MB-231 & Ca Ski cells were either left untreated or treated with metformin or cisplatin alone or metformin in combination with cisplatin for 15 days. Ammonia level in the culture medium was measured as indicated in the Material & Methods section. # Significantly different from treatment with metformin alone. All experiments in this figure were repeated three times. * Significantly different from control untreated cells. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30646605), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - Autophagy & the proteasome regulate CagA stability. (A) Wild-type (WT) & autophagy-deficient (Atg5−/−) MEFs were infected with a CagA+ vacA− isogenic mutant strain (MOI 100) for 8 hours using a gentamycin protection assay. The autophagy marker LC3-II was used to confirm that Atg5−/− MEFs are autophagy-deficient. CagA protein levels were measured by Western blotting using beta -actin as loading control. Graph shows fold change of CagA normalized to beta -actin relative to WT MEFs (mean + SEM; n = 4). (B) AGS cells were infected with a CagA+ vacA− isogenic mutant strain (MOI 50) for 19 hours using a gentamycin protection assay. MG132 (5 μM) was added during the 14-hour low-dose gentamycin incubation. DMSO was used as a vehicle control. CagA protein levels were measured by Western blotting using beta -actin as loading control. Graph shows fold change of CagA normalized to beta -actin relative to vehicle control (mean + SEM; n = 6). (C) AGS cells were infected with a CagA+ vacA− isogenic mutant strain (MOI 50) for 24 hours using a gentamycin protection assay. Lactacystin (10 μM) was added during the 19-hour low-dose gentamycin incubation. Water was used as a vehicle control. CagA protein levels were measured by Western blotting using beta -actin as loading control. Graph shows fold change of CagA normalized to beta -actin relative to vehicle control (mean + SEM; n = 5). Relevant gel bands were cropped from the original blots. Dotted line indicates slicing of two regions together from the same blot. Statistical analysis was performed using Student’s t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30631092), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - Vps34 activity is required for maintaining LC3B-ii levels at P10 & mTOR inhibition increases p-Beclin-1 & LC3B-ii levels at P28. (A) Representative Western blot images for actin, DARPP32 & LC3B-i & -ii in striatal slices obtained from P10 or P28 mice, treated with SAR405 (1 μM) or vehicle (Veh; DMSO, 0.1%). (B,C) Quantification of LC3B-ii relative to actin, normalized to vehicle condition at each age. (B) P10: unpaired, two-tailed t-test, t11 = 2.985, p = 0.0124; P28: unpaired, two-tailed t-test, t24 = 1.807, p = 0.0922. P10: Veh: n = 6 slices, SAR405 n = 7 slices from 3 to 4 mice/age. (C) P28: Veh: n = 9 slices, Baf n = 7 slices from 3 to 4 mice/age. *p 0.05 versus untreated group). C. Morphological features of NB4 cells that were treated with ATRA (1 μM) or HMGB1 (1 μg/ml) was determined by Wright-Giemsa staining & visualized by light microscopy (100X magnification). D. Differentiation of NB4 cells treated with ATRA (1 μM) or HMGB1 (10 μg/ml) for 6-48 h was examined by NBT reduction & surface CD11b expression in comparison to the control (DMSO) (n=3, *P<0.05 versus ATRA group). E. Expression of PML-RAR alpha , P62 & LC3-I/II in NB4 cells treated with ATRA (1 μM) or HMGB1 (10 μg/ml) for 48 h was assayed by western blotting & co-immunoprecipitation in comparison to control (DMSO). Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.15432), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - Zearalenone (ZEN) induced endoplasmic reticulum (ER) stress & autophagy in HepG2 cells. mRNA level & protein expression were determined by quantitative real-time polymerase chain reaction (qRT-PCR) & Western blot analysis, respectively. (A) The protein expression level of GRP78 was measured in cells treated with ZEN for 1 h. (B) The phosphorylation of eIF2 alpha was evaluated in cells treated with ZEN for 2 h. (C) CHOP mRNA levels, (D) Beclin1 mRNA levels, & (E) LC3-II mRNA levels were determined in cells treated with ZEN for 4 h. (F) The protein expression level of LC3-II/LC3-I was examined in cells treated with ZEN for 24 h. Data represent mean ± SEM of three independent experiments. * indicates significant difference vs. the control (** p < 0.01, & *** p < 0.001). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31861425), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - ATRA induces autophagy in SKBR3 cells. (a) SKBR3 & MDA-MB-453 breast cancer cells were treated either with 0.1 or 1 μM ATRA for 2 & 4 days, respectively. beta -Catenin & LC3B levels were measured by western blotting. GAPDH was used as a loading control. (b) LC3B-II quantification from at least three independent experiments. LC3B-II expression was normalized to GAPDH & to vehicle-treated SKBR3 cells. (c) beta -Catenin staining of SKBR3 & MDA-MB453 cells. Cells were control or ATRA (1 μM) treated for 2 days. beta -Catenin FITC (fluorescein isothiocyanate) & nuclear DAPI (4',6-diamidino-2-phenylindole) staining as analyzed by confocal microscopy are shown. (d) Quantification of endogenous LC3B puncta. LC3B in SKBR3 & MDA-MB453 cells treated with 1 μM ATRA for 2 days. (e) LC3B-II western blotting & quantification of the autophagic activation upon treatment with 1 μM ATRA for 2 days in the presence or absence of BafA for 2 h at 200 nM. LC3B-II levels were normalized to GAPDH & to vehicle-treated SKBR3 cells. Standard deviations for five independent experiments are shown. (f) Long-lived protein degradation assay of SKBR3 & MDA-MB453 cells treated as in (e). Radioactivity was determined by liquid scintillation counting of at least three independent experiments. Absolute proteolysis is shown. (g) Long-lived protein degradation assay of SKBR3 treated as in (e) & including treatment with the macroautophagy-specific inhibitor 3-MA (10 mM). Analysis as in (f). Mann–Whitney U-test: *P<0.05, **P<0.01, ***P<0.001 & ****P<0.0001 Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2015236), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - CA1 Tg mice undergo mitochondrial stress. a WB analysis of extracts from 6-mo Wt & Tg mice CA1 (Wt, n = 6; Tg, n = 7), using antibodies against SIRT3, SOD2 & acetylated SOD2. b WB analysis of extracts from 12-mo Wt & Tg mice hippocampi (Wt, n = 5; Tg, n = 5). Data are presented as mean ± SEM (**P < 0.01; ***P < 0.001). WB analysis of extracts from 6-mo Wt & Tg mice CA1 (Wt, n = 6; Tg, n = 5) (c), & from 12-mo Wt & Tg mice hippocampi (Wt, n = 5; Tg, n = 5) (d) for PINK1 & LC-3. Data are presented as mean ± SEM, *P < 0.05. WB analysis of extracts from 6-mo WT & Tg mice CA1 (WT, n = 5; Tg, n = 6) (e), & from 12-mo WT & Tg mice hippocampi (WT, n = 5; Tg, n = 5) (f) for TFAM. Data are presented as mean ± SEM (*P < 0.05), **P < 0.01). Actin was used as loading control Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32131898), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - Bmi‐1 deficiency aggravates cardiac hypertrophy & decreases autophagic flux. (A) Representative micrographs of paraffin‐embedded heart ventricular wall sections of 6‐week‐old Bmi‐1–/– mice & WT mice stained for haematoxylin‐eosin (H&E), wheat germ agglutinin (WGA) & Masson's trichrome (Masson) staining. (B) Heart weight relative to body weight (HW/BW) & heart weight relative to tibia length (HW/TL). (C) The wall thickness of interventricular septum. (D) Myocyte cross‐sectional area from Bmi‐1–/– mice relative to WT mice. (E) Collagenous fibre area from Bmi‐1–/– mice relative to WT mice was counted with anterior wall staining for Masson. (F) Anp, Bnp, beta ‐MHC, c‐TnI & GATA4 mRNA levels in hearts by real‐time RT‐PCR, calculated as ratio to beta ‐actin mRNA & expressed relative to WT. (G) Western blots of cardiac tissue extracts & mouse embryonic cardiomyocytes (MECs) showing ANP, BNP, GATA4, LC3B & p16; beta ‐actin was the loading control. (H) Protein levels relative to beta ‐actin were assessed by densitometric analysis. (I) Representative micrographs of paraffin‐embedded heart sections immunohistochemical staining for ANP, BNP & GATA4. (J) The percentage of cells positive for ANP, BNP, GATA4 or positive area relative to total cells or area. (K) Representative micrographs of fluorescence with autophagy lentivirus transfection, with DAPI for nuclei. (L) Percentage of positive GFP dots or mRFP dots relative to total cells. (M) The number of autophagosomes & autolysosomes per cell. Six mice per group were used for experiments. Cell experiments were performed with three biological repetitions per group. Values are mean ± SEM from six determinations per group, *p < .05, **p < .01, ***p < .001 compared with WT group, unpaired Student's t‐test Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35390228), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - 8-Cl-Ado-induces autophagy. Immunoblot analysis of (A) phospho-ULK (Ser555) levels & (B) LC3B-I lipidation to form LC3B-II in MCF-7 & BT-474 cells treated with 10 μM of 8-Cl-Ado for the indicated times. (C) Fluorescent microscopy of MCF-7 & BT-474 cells transiently transfected with GFP-LC3B & treated with 10 μM of 8-Cl-Ado for 12-hours to assess aggregation of LC3B. (D) Immunoblot analysis of p62 levels in MCF-7 & BT-474 cells treated with 10 μM of 8-Cl-Ado for the indicated times. The normalized ratio of p62 to GAPDH relative to the untreated control is indicated beneath each lane. (E) Fluorescent microscopic imaging of autolysosomes. MCF-7 cells were untreated or treated for 2-days with 8-Cl-Ado or rapamycin. Cells were stained with MDC, blue, to visualize AVO & Syto 61 (DNA), red, for nuclei counterstaining. (F) Representative flow cytometery histograms of AVO stained with acridine orange in MCF-7 cells untreated or treated for 3 days with 50 nM rapamycin or 10 μM 8-Cl-Ado. Baf was added before 30 min prior to staining to neutralize AVO staining. (G) Quantification triplicate experiments of MCF-7 & BT-474 cells treated & analyzed as in E. Image collected & cropped by CiteAb from the following publication (//jhoonline.biomedcentral.com/articles/10.1186/1756-8722-7-23), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3B Antibody - BSA Free [NB600-1384] - PB induces autophagy but blocks autophagosome trafficking & autophagosome-lysosome fusion in Hep3B cells.(A) Hep3B cells, HepG2 cells, & primary mouse hepatocytes treated w/ increasing concentrations of PB (6–2000 μg/mL) for 24 h; viable cells quantified using the MTT assay. Data are presented as mean ± SD from at least 3 independent experiments. Bars = SD. *P < 0.05, **P < 0.01, ***P < 0.001, control versus PB-treated cells. (B) Hep3B cells treated for 6, 24 & 48 h w/ indicated concentrations of PB. Levels of protein expression analysed by WB using antibodies against LC3B, p62/SQSTM1, ATG12, BECN1 & Actin. Images cropped from different blots run under the same experimental conditions. The original blots attached as Supplementary Figure 3A. (C) Hep3B cells transfected w/ GFP-LC3B treated w/ 240 μg/mL PB for 24 h & then observed under a confocal microscope. (D) Hep3B cells treated for 24 h w/ indicated concentrations of PB. Levels of protein expression analysed by WB using antibodies against phosphorylated & total mTOR, 4EBP1, & p70S6K1 as well as actin. Images cropped from different blots run under the same experimental conditions. The original blots attached as Supplementary Figure 3B. (E) Hep3B cells serum-starved overnight, & incubated w/ or w/out PB (240 μg/mL) for 6 h before stimulating w/ 20 ng/mL EGF for 20 min. EGFR protein levels analysed by confocal microscope. (F) Hep3B cells treated PB for 12, 24 or 48 h w/ indicated concentrations. Levels of protein expression analysed by WB using antibodies against EGFR & Actin. Images cropped from different blots run under the same experimental conditions in each panel. The original blots attached as Supplementary Figure 3C. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep41437), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ba, Bv, Ca, Pm, Ye, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ba, Bv, Ca, Pm, Ye, 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	NB600-1384AF350
Alexa Fluor 405	NB600-1384AF405
Alexa Fluor 488	NB600-1384AF488
Alexa Fluor 532	NB600-1384AF532
Alexa Fluor 594	NB600-1384AF594
Alexa Fluor 647	NB600-1384AF647
Alexa Fluor 700	NB600-1384AF700
Alexa Fluor 750	NB600-1384AF750
Allophycocyanin	NB600-1384APC
Biotin	NB600-1384B
CoraFluor 1	NB600-1384CL1
DyLight 350	NB600-1384UV
DyLight 405	NB600-1384V
DyLight 488	NB600-1384G
DyLight 550	NB600-1384R
DyLight 594	NB600-1384DL594
DyLight 650	NB600-1384C
DyLight 680	NB600-1384FR
DyLight 755	NB600-1384IR
FITC	NB600-1384F
HRP	NB600-1384H
Janelia Fluor 525	NB600-1384JF525
Janelia Fluor 549	NB600-1384JF549
Janelia Fluor 646	NB600-1384JF646
Janelia Fluor 669	NB600-1384JF669
PE	NB600-1384PE
PerCP	NB600-1384PCP
mFluor Violet 450 SE	NB600-1384MFV450
mFluor Violet 500 SE	NB600-1384MFV500
mFluor Violet 610 SE	NB600-1384MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-1384SS

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NBP1-49461

Stathmin-2/STMN2 Antibody - BSA Free

(2 Reviews)

(102 Publications)

0.1 mg

NBP1-49461

<b>Genetic Strategies Validation. </b>Staining of STMN2 in primary mouse dorsal root ganglia (DRG) neurons Image shows the expected staining of endogenous STMN2 in axons and growth cones. Tubulin is a marker of axons. NBP1-49461 was used at a dilution of 1:4000. Image courtesy of Dr. Jung Eun Shin.

Neuronal deletion of Tsc2 improves axon regeneration within the first 2 d following nerve injury. SCG10 immunostaining of sciatic nerves 1 d after crush injury in control and Tsc2 cKO mice with (conditioned) or without (sham) a conditioning injury. Red dotted line denotes the crush site while red arrowheads point to the three longest axons. Scale bar: 500 i1/4m. Image collected and cropped by Citeab from the following publication (Nociceptor Deletion of Tsc2 Enhances Axon Regeneration by Inducing a Conditioning Injury Response in Dorsal Root Ganglia. Eneuro (2019) licensed under a CC-BY license.

<b>Expression of Tagged Proteins Validation. </b>Staining of STMN2 in HEK293T cells transfected with GFP or a GFP-STMN2 fusion. NBP1-49461 was used at a dilution of 1:1000 (A, B) and 1:3000 (C). Image courtesy of Dr. Jung Eun Shin.

<b>Expression of Tagged Proteins Validation. </b>Analysis of STMN2 in HEK293T cells transfected with GFP or a GFP-STMN2 fusion. NBP1-49461 was used at a dilution of 1:4000. Image courtesy of Dr. Jung Eun Shin.

Reactivity:

Hu, Mu, Rt, Bv

Species Glossary

Reactivity: Hu, Mu, Rt, Bv

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-49461AF350
Alexa Fluor 405	NBP1-49461AF405
Alexa Fluor 488	NBP1-49461AF488
Alexa Fluor 532	NBP1-49461AF532
Alexa Fluor 594	NBP1-49461AF594
Alexa Fluor 647	NBP1-49461AF647
Alexa Fluor 700	NBP1-49461AF700
Alexa Fluor 750	NBP1-49461AF750
Allophycocyanin	NBP1-49461APC
Biotin	NBP1-49461B
CoraFluor 1	NBP1-49461CL1
DyLight 350	NBP1-49461UV
DyLight 405	NBP1-49461V
DyLight 488	NBP1-49461G
DyLight 550	NBP1-49461R
DyLight 594	NBP1-49461DL594
DyLight 650	NBP1-49461C
DyLight 680	NBP1-49461FR
DyLight 755	NBP1-49461IR
FITC	NBP1-49461F
HRP	NBP1-49461H
Janelia Fluor 525	NBP1-49461JF525
Janelia Fluor 549	NBP1-49461JF549
Janelia Fluor 646	NBP1-49461JF646
Janelia Fluor 669	NBP1-49461JF669
PE	NBP1-49461PE
PerCP	NBP1-49461PCP
mFluor Violet 450 SE	NBP1-49461MFV450
mFluor Violet 500 SE	NBP1-49461MFV500
mFluor Violet 610 SE	NBP1-49461MFV610

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NB600-1335

GADD153/CHOP Antibody (9C8) - BSA Free

(3 Reviews)

(48 Publications)

0.1 ml

NB600-1335

Flow Cytometry: GADD153/CHOP Antibody (9C8) [NB600-1335] - An intracellular stain was performed on SK-MEL-28 cells with GADD153/CHOP Antibody (9C8) NB600-1335 (blue) and a matched isotype control MAB004 (orange). 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 (84540, Thermo Fisher).

<b>Biological Strategies Validation. </b>Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335] - GADD153/CHOP expression in HeLa cells treated with 2.5 ug/mL tunicamycin for 4 hours (Lane 1) and untreated (Lane 2).

<b>Biological Strategies Validation. </b>Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335] - Analysis of endogenous CHOP/GADD153 from primary human fibroblasts using NB600-1335. Lane 1: Untreated cells, Lane 2: Cells treated with tunicamycin for 10 hours.

Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335] - Analysis of CHOP in rat heart tissue lysate. Image courtesy of product review submitted by Lee Hsiao-Wei.

Simple Western: GADD153/CHOP Antibody (9C8) [NB600-1335] - Image shows a specific band for CHOP/GADD153 in 1.0 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

The TUG1‐miR‐144‐3p‐Nrf2 axis regulates H/R‐induced cell apoptosis by adjusting oxidative stress and endoplasmic reticulum stress in vitro. A, B, TUNEL stain and quantitative analysis for H/R‐injured TCMK cells with different transfections (n = 3). C–J, The expression levels of Nrf2, H0‐1, CHOP, GRP78, Bax, Bcl‐2 and cleaved‐caspase3 in H/R‐injured TCMK cells with different transfections were measured by Western blot analysis. Representative protein bands are shown in F, and the quantitative analysis of protein expression is shown in C‐E, G‐J (n = 3). K, L, The levels of SOD and MDA in different transfection groups (n = 3). All data are expressed as the mean +/- SD; data comparisons between multiple groups were performed using one‐way analysis of variance (ANOVA) with Tukey's post hoc test. *p < 0.05, **p < 0.01 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34547172), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

MiR‐144‐3p regulates H/R‐induced Nrf2‐HO‐1 signaling pathway activation, oxidative stress, mitochondria and endoplasmic reticulum functions in vitro. A–C, The expression levels of Nrf2 and HO‐1 in TCMK cells transfected with miR‐144‐3p mimics or inhibitor after H/R treatment. Representative bands are shown in A, and the quantitative analysis of protein expression level is shown in B, C (n = 3). D, E, The SOD and MDA levels in cellular supernatant of TCMK cells transfected with miR‐144‐3p mimics or inhibitor after H/R treatment (n = 3). F–I, The expression levels of CHOP, GRP78 and Cytochrome C in TCMK cells transfected with miR‐144‐3p mimics or inhibitor after H/R treatment. Representative bands are shown in F, and the quantitative analysis of protein expression level is shown in G–I (n = 3). All data are expressed as the mean +/- SD; data comparisons between multiple groups were performed using one‐way analysis of variance (ANOVA) with Tukey's post hoc test. *p < 0.05, **p < 0.01 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34547172), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

UPR Western blotting in vitro (A). (B) ASC-CO and ASC-CM reduced the expression of BIP under exposure to CoCl2, incubations with tunicamycin, and CoCl2 were used as controls (C) ASC-CO and ASC-CM reduced the protein expression of ATF6 under CoCl2, incubations with tunicamycin, and CoCl2 were used as controls (D) ASC-CO 57 and ASC-CM reduced the protein expression of XBP1, incubations with tunicamycin, and CoCl2. (E) The protein expression of CHOP and tunicamycin remained unchanged with ASC-CO and ASC-CM as compared to controls. (n = 3, ANOVA ns, not significant; * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001). Image collected and cropped by CiteAb from the following open publication (//www.mdpi.com/1422-0067/24/24/17197), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Knocking down Nrf2 reversed the miR‐144‐3p inhibitor‐alleviated H/R‐induced apoptosis, oxidative stress, mitochondrial damage and endoplasmic reticulum stress in vitro. A–c, F–K, The protein expression level of Nrf2, HO‐1, cleaved‐caspase3, Bax, Bcl‐2, CHOP, GRP78 and Cytochrome C in H/R‐injured TCMK cells transfected with miR‐144‐3p inhibitor or si‐Nrf2. Representative bands are shown in A, and the quantitative analysis of protein expression is shown in B, C, F–K (n = 3). D, E, The SOD and MDA levels in cellular supernatants in H/R‐injured TCMK cells transfected with miR‐144‐3p inhibitor or si‐Nrf2 (n = 3). All data are expressed as the mean +/- SD; data comparisons between multiple groups were performed using one‐way analysis of variance (ANOVA) with Tukey's post hoc test. *p < 0.05, **p < 0.01, NS, no significant difference Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34547172), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

TUG1 plays an important role in H/R‐induced cell apoptosis possibly through regulating the Nrf2‐HO‐1 pathway, oxidative stress, mitochondrial damage and endoplasmic reticulum stress via targeting miR‐144‐3p. A, The expression of miR‐144‐3p in TUG1‐overexpressing or TUG1‐knockdown TCMK cells detected by RT‐qPCR (n = 3). B, C, The levels of SOD and MDA in cellular supernatant (n = 3). D–J, The relative expression levels of Nrf2, HO‐1, GRP78, CHOP, Cyt C and cleaved‐caspase3 were examined. Representative protein bands are shown in D, and the quantitative analysis of protein expression is shown in E‐J (n = 3). All data are expressed as the mean +/- SD; data comparisons between multiple groups were performed using one‐way analysis of variance (ANOVA) with Tukey's post hoc test. *p < 0.05, **p < 0.01 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34547172), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

ASCs lead to changes in UPR expression in vivo (A) The comparison of the left and right legs—with DLFA- of the two groups of mice are shown (B) Ratio between BIP expression between left and right leg with and without ASCs (C) Ratio of ATF6 expression between left and right leg with and without ASCs (D) Ratio of CHOP expression between left and right leg with and without ASCs (E) Ratio of XBP1 expression between left and right leg with and without ASCs (n = 3, ANOVA, * p ≤ 0.05, ** p ≤ 0.01). Image collected and cropped by CiteAb from the following open publication (//www.mdpi.com/1422-0067/24/24/17197), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Response to ER Stress Mediates the Reprogramming Facilitation by Surf4. (A) The RNA level of ER stress‐related genes on day 3 of reprogramming with or without exogenous Surf4. Relative expression of these genes relative to beta ‐actin (n = 3, average +/-SEM). (B) The protein level of ER stress‐related genes on day 3 of reprogramming with or without exogenous Surf4. (C) Kinetics of Oct4‐GFP+colony formation with or without exogenous Surf4 and sXbp1‐ delta DBD during reprogramming. (D) The number of Oct4‐GFP+ colonies and the percentage of Oct4‐GFP+ cells induced by OSKM plus Surf4 and sXbp1‐ delta DBD. (E) Morphology of the primary colonies induced by OSKM plus Surf4 and sXbp1‐ delta DBD. Scale bars, 1000 μm. Magnification: ×40. (F) AP staining of the primary iPS colonies. See also Figure S4 and Table S1 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34585448), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Pm

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-1335AF350
Alexa Fluor 405	NB600-1335AF405
Alexa Fluor 488	NB600-1335AF488
Alexa Fluor 532	NB600-1335AF532
Alexa Fluor 594	NB600-1335AF594
Alexa Fluor 647	NB600-1335AF647
Alexa Fluor 700	NB600-1335AF700
Alexa Fluor 750	NB600-1335AF750
Allophycocyanin	NB600-1335APC
Biotin	NB600-1335B
CoraFluor 1	NB600-1335CL1
DyLight 350	NB600-1335UV
DyLight 405	NB600-1335V
DyLight 488	NB600-1335G
DyLight 550	NB600-1335R
DyLight 594	NB600-1335DL594
DyLight 650	NB600-1335C
DyLight 680	NB600-1335FR
DyLight 755	NB600-1335IR
FITC	NB600-1335F
HRP	NB600-1335H
Janelia Fluor 525	NB600-1335JF525
Janelia Fluor 549	NB600-1335JF549
Janelia Fluor 646	NB600-1335JF646
Janelia Fluor 669	NB600-1335JF669
PE	NB600-1335PE
PerCP	NB600-1335PCP
mFluor Violet 450 SE	NB600-1335MFV450
mFluor Violet 500 SE	NB600-1335MFV500
mFluor Violet 610 SE	NB600-1335MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-1335SS
Azide and BSA Free	NBP2-80745

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NB300-130

Ubiquitin Antibody (Ubi-1) - BSA Free

(4 Reviews)

(53 Publications)

0.1 ml

NB300-130

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Analysis of HEK293 cell lysates using mouse mAb Ubiquitin, dilution 1:1000 (Green). [1] protein standard (Red), [2] cells maintained in normal medium, [3] cells treated with 10uM of proteasome inhibitor lactacystin (Lc) for 16hrs. Lysed cells were electrophoresed on 4-20% SDS-PAGE and transferred to PVDF membranes. The smear detected above the 200kDa standard represent accumulation of ubiquitinated proteins in proteasome inhibitor-Lc treated cells. The prominent band at 8kDa corresponds to monoubiquitin. Rabbit pAb to HSP60, dilution 1:5000 (Red) was used as a loading control.

Immunohistochemistry-Paraffin: Ubiquitin Antibody (Ubi-1) [NB300-130] - FFPE section of cerebral cortex of an Alzheimer patient processed with Ubiquitin antibody using HRP/DAB (Brown) and also stained with haemotoxylin (Blue). A typical flame shaped tangle is seen in a pyramidal neuron in the center and is surrounded by some dystrophic neurites, also strongly ubiquitin positive. Both are commonly seen in cortical and hippocampal Alzheimer brain sections and are typical for this disease, but are rare or absent in healthy brain.

Immunohistochemistry-Paraffin: Ubiquitin Antibody (Ubi-1) [NB300-130] - Staining of Ubiquitin in hippocampal tissue from an Alzheimer patient.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Molecular Analysis of XopD Ub/Ubl Specificity. S. lycopersicum protein extract blotted for total Ub (Ubiquitin; Novus Biologicals) following a 1 hr room temperature treatment with the XopD variants used in (K) at 1 uM final concentration. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27425412/) licensed under a CC-BY license.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - Western blot of Ubiquitin expression on (Lane 1) poly-ubiquitin (lys63 linked), (Lane 2) pure ubiquitin and crude homogenates of adult rat, (Lane 3) cortex, (Lane 4) cerebellum and (Lane 5) brain stem.

Immunohistochemistry-Paraffin: Ubiquitin Antibody (Ubi-1) [NB300-130] - IHC staining of Ubiquitin in human rectal cancer using DAB with hematoxylin counterstain.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - In Vivo Applications of Affimers(A) HA-NleL 293 T-Rex cells were induced with 1 μg/mL doxycycline for 12 hr or left untreated. Whole-cell lysate (WCL) blots are shown for actin, HA(-NleL), Ub, & K6 chains. Western blots with the K6 affimer are also shown after Ub enrichment using TUBEs.(B) TUBE-PD of HEK293 cells after 1 hr of MG132 (10 μM) without further treatment (−) or with additional UV (40 J/m2) or CCCP treatment (10 μM for 1 hr) & subsequently blotted with the K6 affimer. Input controls are shown for total Ub & actin & gamma H2AX. The relative signal increase from two experiments is shown below the respective lanes.(C) Expression of WT or catalytically inactive (C431S) Parkin in HeLa Flp-In cells was induced with 0.2 μg/mL doxycycline for 16 hr. Mitochondria were depolarized with O/A for 2 hr. WCL inputs are shown for total Ub, expressed Parkin, TOM20, & actin. The TUBE-PD was also blotted using the K6 affimer & total Ub.(D) Confocal fluorescence microscopy images of cells as in (B) stained with K6 affimer (green), TOM20 (red), & DAPI (blue). Cells & a magnified area are outlined in white. Scale bars correspond to 20 μm.See also Figure S5. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28943312), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ubiquitin Antibody (Ubi-1) [NB300-130] - HUWE1 Assembles K6 Chains In Vitro & In Vivo(A) Table summarizing proteins identified with the corresponding number peptide-spectrum matches (PSMs) in three replicates of DUB-treated K6 affimer pull-downs.(B) In vitro assembly reaction of the HECT E3 HUWE1 with Ub WT, Ub K6R, Ub K11R, & Ub K48R on Coomassie. Arrows indicate K6 diUb.(C) Linkage composition of HUWE1-generated diUb after 1 hr as determined by AQUA MS.(D) AQUA-derived total cellular chain composition of HUWE1−/− HeLa & parental cells after TUBE-based enrichment. Error bars indicate mean ± SD from n = 3. ∗p < 0.05, according to a two-tailed Student’s t test. N.S., not significant.(E) TUBE-PD from a doxycycline-inducible HUWE1 shRNA Ls174T cell line blotted with the K6-specific affimer, with input controls for actin, total Ub, & HUWE1.(F) K6 affimer pull-down in doxycycline-inducible HUWE1 shRNA Ls174T cells blotted against Mfn2. Cells were left untreated or treated with 10 μg/mL MG132 for 4 hr and/or 1 μg/mL doxycycline for 72 hr. Pull-downs were incubated with 250 nM USP21 as indicated, with a K48 blot to show completeness of the deubiquitination reaction. Input controls are shown for Mfn2, actin, & HUWE1.See also Figure S7. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28943312), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ce, Ch, Dr, Eq, Pl, Pm, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ce, Ch, Dr, Eq, Pl, Pm, Ze

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	NB300-130AF350
Alexa Fluor 405	NB300-130AF405
Alexa Fluor 488	NB300-130AF488
Alexa Fluor 532	NB300-130AF532
Alexa Fluor 594	NB300-130AF594
Alexa Fluor 647	NB300-130AF647
Alexa Fluor 700	NB300-130AF700
Alexa Fluor 750	NB300-130AF750
Allophycocyanin	NB300-130APC
Biotin	NB300-130B
CoraFluor 1	NB300-130CL1
DyLight 350	NB300-130UV
DyLight 405	NB300-130V
DyLight 488	NB300-130G
DyLight 550	NB300-130R
DyLight 594	NB300-130DL594
DyLight 650	NB300-130C
DyLight 680	NB300-130FR
DyLight 755	NB300-130IR
FITC	NB300-130F
HRP	NB300-130H
Janelia Fluor 525	NB300-130JF525
Janelia Fluor 549	NB300-130JF549
Janelia Fluor 646	NB300-130JF646
Janelia Fluor 669	NB300-130JF669
PE	NB300-130PE
PerCP	NB300-130PCP
mFluor Violet 450 SE	NB300-130MFV450
mFluor Violet 500 SE	NB300-130MFV500
mFluor Violet 610 SE	NB300-130MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB300-130SS

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NB100-1612

beta-III Tubulin Antibody - BSA Free

(2 Reviews)

(55 Publications)

0.25 ml

NB100-1612

beta-III Tubulin Western Blot using homogenates of adult mouse brain (1:3000).

Western Blot: beta-III Tubulin Antibody [NB100-1612] - The levels of glial fibrillary acidic protein (GFAP) & S100 beta as astrocytic protein marker proteins in the hippocampal area. (A) Western blot images using GFAP, tubulin beta‐3, & beta ‐actin antibodies to total protein lysate in hippocampal area. The statistical results of internal standardized GFAP/ beta ‐tubulin ratio by beta ‐actin levels relative to young adult wild‐type C57BL/6J. White & black bars indicate the wild‐type C57BL/6J & Tet‐mev‐1 mice, respectively. Data are expressed as mean ± SD; *P 12 in each group. (B) Western blot images using S100 beta , tubulin beta‐3, & beta ‐actin antibodies to total protein lysate in hippocampal area. The statistical results of internal standardized S100 beta / beta ‐tubulin ratio by beta ‐actin levels relative to young adult wild‐type C57BL/6J. White & black bars indicate the wild‐type C57BL/6J & Tet‐mev‐1 mice, respectively. Data are expressed as mean ± SD; *P 12 in each group. (C) Micrographs of immunohistochemical analysis on paraffined hippocampal tissue sections using GFAP & S100 beta antibody. Brown cells indicate GFAP‐stained astrocytes. Scale bar = 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27623715), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , ,

Applications: , , , , ,

Host:

Chicken Polyclonal

Host: Chicken Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-1612-0.05ml

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NB100-402

SLC31A1/CTR1 Antibody - BSA Free

(37 Publications)

0.1 ml

NB100-402

Immunohistochemistry: SLC31A1/CTR1 Antibody [NB100-402] - Left panel: Antibody staining of hCTR1 overexpressing cell line. Right panel: Antibody plus peptide competition staining.

SLC31A1/CRT1 was detected in immersion fixed NIH3T3 Mouse fibroblast cell line using Rabbit anti-SLC31A1/CRT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # NB100-402) at 2 µg/mL overnight at 4C. Cells were stained using DyLight 488-conjugated Anti-Rabbit IgG (H+L) Cross-Absorbed Secondary Antibody (green) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Identification of the copper transporter CTR1 in porcine oocytes and cumulus cells. (A) Immunofluorescence indicating the localization of the CTR1 in meiosis II stage-oocytes and mature cumulus cells. Scale bar = 50 and 20 μm, respectively. (B) A representative western blot image and relative densitometric bar graph of the CTR1 expression in granulosa cells, immature cumulus, mature cumulus, meiosis I (MI) oocytes, and meiosis II (MII) oocytes. An asterisk indicates statistical significance (*p < 0.05). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36158185), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ca, Xp, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca, Xp, Ze

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	NB100-402AF350
Alexa Fluor 405	NB100-402AF405
Alexa Fluor 488	NB100-402AF488
Alexa Fluor 532	NB100-402AF532
Alexa Fluor 594	NB100-402AF594
Alexa Fluor 647	NB100-402AF647
Alexa Fluor 700	NB100-402AF700
Alexa Fluor 750	NB100-402AF750
Allophycocyanin	NB100-402APC
Biotin	NB100-402B
CoraFluor 1	NB100-402CL1
DyLight 350	NB100-402UV
DyLight 405	NB100-402V
DyLight 488	NB100-402G
DyLight 550	NB100-402R
DyLight 594	NB100-402DL594
DyLight 650	NB100-402C
DyLight 680	NB100-402FR
DyLight 755	NB100-402IR
FITC	NB100-402F
HRP	NB100-402H
Janelia Fluor 525	NB100-402JF525
Janelia Fluor 549	NB100-402JF549
Janelia Fluor 646	NB100-402JF646
Janelia Fluor 669	NB100-402JF669
PE	NB100-402PE
PerCP	NB100-402PCP
mFluor Violet 450 SE	NB100-402MFV450
mFluor Violet 500 SE	NB100-402MFV500
mFluor Violet 610 SE	NB100-402MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-402SS

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NBP2-13172

GADD153/CHOP Antibody - BSA Free

(1 Review)

(27 Publications)

0.1 ml

NBP2-13172

<b>Biological Strategies Validation. </b>Western Blot: GADD153/CHOP Antibody - BSA Free [NBP2-13172] - GADD153/CHOP Antibody [NBP2-13172] - Effects of psoralen on ER-stress related protein expression in SMMC7721. Western blot assays showing the effects of psoralen and thapsigargin on ER-stress related protein expression. Image collected and cropped by CiteAb from the following publication (//biolres.biomedcentral.com/articles/10.1186/s40659-019-0241-8), licensed under a CC-BY license.

Immunohistochemistry: GADD153/CHOP Antibody [NBP2-13172] - Mouse liver tissue. Antigen retreival: Citrate buffer pH=6.0,Blocking: 10% BSA in 0.1%Tween in TBS, Primary antibody: 1:100 in 1% BSA in 0.1%Tween in TBS,Incubation: Overnight at 4C, Secondary antibody: 1:200 incubation for 1hr at room temperature, AB complex: 1:100 for 1hr at room temprature Develpment DAB. IHC-P image submitted by a verified customer review.

Flow Cytometry: GADD153/CHOP Antibody [NBP2-13172] - An intracellular stain was performed on SK-MEL-28 cells with GADD153/CHOP Antibody NBP2-13172F (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 FITC.

<b>Biological Strategies Validation. </b>Western Blot: GADD153/CHOP Antibody [NBP2-13172] - CHOP/GADD153 Antibody [NBP2-13172] - WB analysis of CHOP/GADD153 in tunicamycin treated (+) and untreated (-) HeLa cell lysate.

Immunohistochemistry: GADD153/CHOP Antibody [NBP2-13172] - IHC staining of CHOP/GADD153 in mouse colon using DAB with hematoxylin counterstain.

Western Blot: GADD153/CHOP Antibody - BSA Free [NBP2-13172] - Effects of psoralen on ER-stress related protein expression in SMMC7721. a Western blot assays showing the effects of psoralen & thapsigargin on ER-stress related protein expression. b Western blot assays showing the effects of TUDC on the expression of ER-stress related protein induced by PSO. c The mRNA levels of GRP78 & DDIT3 with TUDC under PSO & TG treatment. Values are mean ± SD (n = 3) & * is means compared to the Con group, **P < 0.01 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31277690), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GADD153/CHOP Antibody - BSA Free [NBP2-13172] - Reduction of fibronectin prevents Dex-induced ER stress in human GTM-3 cells. (A) GTM3 cells were transfected with a plasmid expressing CRISPR-Cas9 targeting fibronectin (CR-FN) & then treated with Veh or Dex for 48 hours. Fibronectin knockdown partially reduced ER stress as evident from reduced GRP78 & CHOP levels after Dex treatment (n = 2). (B) Human GTM3 cells were treated with Dex (100 nM) with or without CR-FN for 48 hours. Fixed cells were stained for fibronectin & KDEL to examine Dex-induced fibronectin & ER stress. Fibronectin knockdown reduced fibronectin & ER stress in Dex-treated TM cells (n = 2). Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-14938-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GADD153/CHOP Antibody - BSA Free [NBP2-13172] - TM cells are sensitive to ER stress from Dex-induced abnormal ECM compared to other ocular cells: Primary human TM cells (A), RPE (B) & corneal fibroblasts (C) were treated with Veh or Dex for 3 days & immunostained for fibronectin & KDEL. Although fibronectin was increased in all of these cell types, ER stress was only observed in TM cells. (D) Human GTM3 or ARPE-19 cells were treated with Veh or Dex for 3 days. Fibronectin & chronic ER stress markers ATF4 & CHOP were examined in cellular lysates (n = 2). Although Dex increased fibronectin in both GTM3 & ARPE-19 cells, chronic ER stress was only observed in GTM3 cells indicating that TM cells are more prone to ER stress from Dex-induced ECM accumulation. (E) GTM-3 or ARPE-19 cells were treated with cellular fibronectin (cFn) for 2 days & ER stress was examined by Western blot analysis. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-14938-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GADD153/CHOP Antibody - BSA Free [NBP2-13172] - Overexpression of extracellular fibronectin induces ER stress in primary human TM cells. (A) Primary human TM cells (n = 2 strains) were treated with cellular fibronectin (10 ug/ml) for 48 hours & cellular lysates were analyzed for ER stress markers. (B) Primary human TM cells (n = 2 strains) were treated with cellular fibronectin (10 ug/ml) for 48 hours & fixed cells were stained with fibronectin & KDEL antibodies. Cellular fibronectin increased extracellular fibronectin & ER stress in GTM3 cells (n = 2). Scale bar = 50 μm. (C) GTM3 cells were transduced with fibronectin activation lentiviral particles for 48 hours & stained with fibronectin & KDEL antibodies. Increased fibronectin expression alone is sufficient to induce ER stress in TM cells (n = 2). Scale bar = 50 μm. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-14938-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GADD153/CHOP Antibody - BSA Free [NBP2-13172] - NASTRp induces ER stress & eventually leads to cell death with Bim up-regulation.(A) qRT-PCR analysis of biomarkers of ER stress/UPR related genes in NCI-H441 treated with 20 μM NASTRp in time course manner. (B) NASTRp induced ER stress & activated UPR led to apoptosis with Bim induction in NSCLC cell lines. Cells were treated with the indicated concentrations of NASTRp for 24 hours & cell lysates were subjected to SDS-PAGE/Western blot analysis using the indicated antibodies. (C) Upregulation of CHOP by NASTRp. NCI-H441 cells were treated with 20 NASTRp for 0–24 hours. At the indicated time points, cells were fixed & subjected to immunofluorescence assay with anti-CHOP antibody. Cell images were microphotographed using fluorescence microscopy at 60X magnification. Scale bars: white; 10 μm. (D) TUNEL-positive cells in NASTRp-treated (for 48 hours) A549 cells were counted & analyzed as relative % of TUNEL-positive cells. *P < 0.05 versus vehicle. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0122628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: GADD153/CHOP Antibody - BSA Free [NBP2-13172] - NASTRp induces ER stress & eventually leads to cell death with Bim up-regulation.(A) qRT-PCR analysis of biomarkers of ER stress/UPR related genes in NCI-H441 treated with 20 μM NASTRp in time course manner. (B) NASTRp induced ER stress & activated UPR led to apoptosis with Bim induction in NSCLC cell lines. Cells were treated with the indicated concentrations of NASTRp for 24 hours & cell lysates were subjected to SDS-PAGE/Western blot analysis using the indicated antibodies. (C) Upregulation of CHOP by NASTRp. NCI-H441 cells were treated with 20 NASTRp for 0–24 hours. At the indicated time points, cells were fixed & subjected to immunofluorescence assay with anti-CHOP antibody. Cell images were microphotographed using fluorescence microscopy at 60X magnification. Scale bars: white; 10 μm. (D) TUNEL-positive cells in NASTRp-treated (for 48 hours) A549 cells were counted & analyzed as relative % of TUNEL-positive cells. *P < 0.05 versus vehicle. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0122628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, 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	NBP2-13172AF350
Alexa Fluor 405	NBP2-13172AF405
Alexa Fluor 488	NBP2-13172AF488
Alexa Fluor 532	NBP2-13172AF532
Alexa Fluor 594	NBP2-13172AF594
Alexa Fluor 647	NBP2-13172AF647
Alexa Fluor 700	NBP2-13172AF700
Alexa Fluor 750	NBP2-13172AF750
Allophycocyanin	NBP2-13172APC
Biotin	NBP2-13172B
CoraFluor 1	NBP2-13172CL1
DyLight 350	NBP2-13172UV
DyLight 405	NBP2-13172V
DyLight 488	NBP2-13172G
DyLight 550	NBP2-13172R
DyLight 594	NBP2-13172DL594
DyLight 650	NBP2-13172C
DyLight 680	NBP2-13172FR
DyLight 755	NBP2-13172IR
FITC	NBP2-13172F
HRP	NBP2-13172H
Janelia Fluor 525	NBP2-13172JF525
Janelia Fluor 549	NBP2-13172JF549
Janelia Fluor 646	NBP2-13172JF646
Janelia Fluor 669	NBP2-13172JF669
PE	NBP2-13172PE
PerCP	NBP2-13172PCP
mFluor Violet 450 SE	NBP2-13172MFV450
mFluor Violet 500 SE	NBP2-13172MFV500
mFluor Violet 610 SE	NBP2-13172MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-13172-0.025ml

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NB300-135

NF-H Antibody

(30 Publications)

0.05 ml

NB300-135

Immunohistochemistry: NF-H Antibody [NB300-135] - Immunostainings of adult organotypic brain slices at the hippocampal level. Brain slices were prepared from WT or TG adult mice and incubated with okadaic acid (OA). Brain slices were stained for nuclear DAPI (blue, A,B) showing intact brain structures. Thiazine Red staining (C,D) shows red fluorescent plaques in the hippocampus (D) of TG mice, while only background is seen in the hippocampus (C) of WT mice. Staining of neurofilament fibers (G,H) shows good neuronal viability both in WT and TG mice. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fnagi.2018.00113/full) licensed under a CC-BY license.

Western Blot: NF-H Antibody [NB300-135] - Analysis of 200kDa Neurofilament Heavy expression in rat spinal cord extract. The first lane is Coomassie Brilliant Blue stained and the second lane is probed with rabbit anti-Neurofilament Heavy antibody NB300-135. The NF-H corresponds to a weight of 200kDa.

Western Blot: NF-H Antibody [NB300-135] - Different tissue lysates using rabbit pAb to NF-H, dilution 1:10,000 in green: [1] protein standard (red), [2] rat brain, [3] rat spinal cord [4] mouse brain, and [5] mouse spinal cord lysate. Strong band at about 220 kDa corresponds to the phosphorylated axonal form of the NF-H subunit. Smaller proteolytic fragments of NF-H are also detected with RPCA-NF-H antibody.

Reactivity:

Hu, Mu, Rt, Po, Bv, Eq

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Eq

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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NB100-513

USP7 Antibody - BSA Free

(1 Review)

(30 Publications)

100 ul

NB100-513

Immunohistochemistry-Paraffin: USP7 Antibody [NB100-513] - Sample: FFPE section of mouse renal cell carcinoma.Antibody: Affinity purified rabbit anti-USP7 used at 1:5,000 (0.2ug/ml). Detection:DAB

Immunoprecipitation: USP7 Antibody [NB100-513] - Samples: Wholecell lysate (10 ug) from HEK293T, RKO, Hep-G2, Jurkat,HeLa, TCMK-1, and NIH 3T3 cells prepared using NETNlysis buffer. Antibody: Affinity purified rabbit anti-USP7antibody used for WB at0.04 ug/ml. Detection: Chemiluminescence with anexposure time of 10 seconds

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-513AF350
Alexa Fluor 405	NB100-513AF405
Alexa Fluor 488	NB100-513AF488
Alexa Fluor 532	NB100-513AF532
Alexa Fluor 594	NB100-513AF594
Alexa Fluor 647	NB100-513AF647
Alexa Fluor 700	NB100-513AF700
Alexa Fluor 750	NB100-513AF750
Allophycocyanin	NB100-513APC
Biotin	NB100-513B
CoraFluor 1	NB100-513CL1
DyLight 350	NB100-513UV
DyLight 405	NB100-513V
DyLight 488	NB100-513G
DyLight 550	NB100-513R
DyLight 594	NB100-513DL594
DyLight 650	NB100-513C
DyLight 680	NB100-513FR
DyLight 755	NB100-513IR
FITC	NB100-513F
HRP	NB100-513H
Janelia Fluor 525	NB100-513JF525
Janelia Fluor 549	NB100-513JF549
Janelia Fluor 646	NB100-513JF646
Janelia Fluor 669	NB100-513JF669
PE	NB100-513PE
PerCP	NB100-513PCP
mFluor Violet 450 SE	NB100-513MFV450
mFluor Violet 500 SE	NB100-513MFV500
mFluor Violet 610 SE	NB100-513MFV610

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NBP1-78444

Mas Antibody - BSA Free

(2 Reviews)

(21 Publications)

0.1 ml

NBP1-78444

Immunohistochemistry: Mas Antibody [NBP1-78444] - Analysis of MAS1 in mouse kidney using DAB with hematoxylin counterstain.

Simple Western: Mas Antibody [NBP1-78444] - Lane view shows a specific band for MAS1 in 0.5 mg/ml of HepG2 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

MasR activation restores LPS‐induced overactivation of ACE/AngII/AT1 axis and contains neuroprotective properties. (a–j) The impacts of MasR agonist, AVE, and the ACE2 activator, DIZE, on brain RAS following repeated LPS treatment. (a) Representative western blots. ACE protein expression (b) and activity (c). (d) AngII concentration. (e) AT1 protein expression. ACE2 protein expression (f) and activity (g). (h) Ang(1–7) concentration. MasR protein expression (i) and immunofluorescence staining (j). (m) Representative images of TUNEL and Nissl staining. (k, l) mRNA expression of biomarkers of microglial M1 phenotype (k) and M2 phenotype (l). Scale bar = 50 μm. Data are means +/- SD (n = 7). **p < 0.01 compared to control group. +p < 0.05, ++p < 0.01 compared to LPS group Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34529881), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

LPS exposure shifts the balance of brain RAS. (a) The impacts of acute LPS (AcLPS) or repeated LPS (ReLPS) exposure on microglial activation (IBA‐1 staining) and ROS generation (DHE staining) in the brain cortex of C57BL/6 mice. (b) Biomarkers of microglial M1 phenotype mRNA expression. (c) Biomarkers of microglial M2 phenotype mRNA expression. (d–m) Alterations of ACE/AngII/AT1 and ACE2/Ang(1–7)/MasR pathways following LPS treatment. (d) Representative western blots. ACE protein expression (e) and activity (f). (g) AngII concentration. (h) AT1 protein expression. (i) AT2 protein expression. ACE2 protein expression (j) and activity (k). (L) Ang(1–7) concentration. (m) MasR protein expression. Scale bar = 50 μm. Data are means +/- SD (n = 7–9). *p < 0.05, **p < 0.01 compared to control group Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34529881), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Pm, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, 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 350	NBP1-78444AF350
Alexa Fluor 405	NBP1-78444AF405
Alexa Fluor 488	NBP1-78444AF488
Alexa Fluor 532	NBP1-78444AF532
Alexa Fluor 594	NBP1-78444AF594
Alexa Fluor 647	NBP1-78444AF647
Alexa Fluor 700	NBP1-78444AF700
Alexa Fluor 750	NBP1-78444AF750
Allophycocyanin	NBP1-78444APC
Biotin	NBP1-78444B
CoraFluor 1	NBP1-78444CL1
DyLight 350	NBP1-78444UV
DyLight 405	NBP1-78444V
DyLight 488	NBP1-78444G
DyLight 550	NBP1-78444R
DyLight 594	NBP1-78444DL594
DyLight 650	NBP1-78444C
DyLight 680	NBP1-78444FR
DyLight 755	NBP1-78444IR
FITC	NBP1-78444F
HRP	NBP1-78444H
Janelia Fluor 525	NBP1-78444JF525
Janelia Fluor 549	NBP1-78444JF549
Janelia Fluor 646	NBP1-78444JF646
Janelia Fluor 669	NBP1-78444JF669
PE	NBP1-78444PE
PerCP	NBP1-78444PCP
mFluor Violet 450 SE	NBP1-78444MFV450
mFluor Violet 500 SE	NBP1-78444MFV500
mFluor Violet 610 SE	NBP1-78444MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP1-78444SS

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NB200-111

p14ARF/CDKN2A Antibody - BSA Free

(2 Reviews)

(19 Publications)

0.1 ml

NB200-111

Immunocytochemistry/Immunofluorescence: p14ARF/CDKN2A Antibody [NB200-111] - p14ARF antibody was tested in HeLa cells with Dylight 488 (green). Nuclei were counterstained with DAPI (blue). Tubulin was stained with alpha tubulin (red).

<b>Biological Strategies Validation. </b>Western Blot: p14ARF/CDKN2A Antibody [NB200-111] - ARF expression levels in OCI/AML3 cells after proteasome treatment. This image was submitted via customer Review.

Flow Cytometry: p14ARF/CDKN2A Antibody [NB200-111] - p14ARF antibody was tested at 1:400 in HeLa cells using an Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.

Western Blot: p14ARF/CDKN2A Antibody [NB200-111] - Analysis of HeLa Whole Cell Lysate (NB800-PC1) using p14ARF antibody (lot C) employing ECL detection method.

Immunohistochemistry-Frozen: p14ARF/CDKN2A Antibody [NB200-111] - Mouse brain section, hippocampal (10 um thickness). 3% PFA perfused. 1:1000 dilultion. Image from verified customer review. Image using the Biotin form of this antibody.

Western Blot: p14ARF/CDKN2A Antibody - BSA Free [NB200-111] - Inhibition of AKT decreases p53mut stability(A) T24 cells & PSN1 cells were treated with MK-2206 (5 μM) or DMSO for 24 hrs. Cells were fixed & stained with DAPI & anti-mutant p53 (OP29 clone). (B) T24 cells were pre-treated with MK-2206 (5 μM) or DMSO for 24 hrs before the addition of fresh media containing cyclohexamide (100 μM) (CHX) in combination with MK-2206 (5 μM) or DMSO for the times indicated. Nuclear extracts were prepared from treated cells & blotted with the indicated antibodies. Quantification is relative to initiation of CHX treatment for both conditions *, MK-2206 is 40% of DMSO control but taken as 1.0 for relative assessment. (C) H1299 cells were transfected with HA-tagged-ubiquitin & mutant p53 (R175H or R248W) as indicated. Transfected cells were treated with DMSO, MK-2206 (5 μM) or Nutlin3A (5 μM) for 16hrs as indicated. p53 immunoprecipitates & whole cell lysates were probed with the indicated antibodies. (D) T24 cells were transfected with non-targeting control or p14ARF siRNA & treated with DMSO, MK-2206 (5 μM) or the NPM oligomerisation inhibitor NCS348884 (4 μM) (Qi et al., 2008) as indicated. (E) T24 cells were transfected with non-targeting control, AKT1, or p14ARF siRNA. Cells were treated with NCS348884 (4 μM), Nutlin3A (5 μM) or DMSO as indicated. Whole cell lysates were probed with the indicated antibodies. (F) KPC mice derived KRASG12D p53 Floxed (p53Fl), KRASG12D p53R172H ARF+/+ & KRASG12D p53R172H ARF−/− pancreatic tumor cells were treated with MK-2206 (1μM), Nutlin3A (5μM) or DMSO as indicated. Whole cell lysates were probed with the indicated antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: p14ARF/CDKN2A Antibody - BSA Free [NB200-111] - Inhibition of AKT modulates p53 stability in-vivo & synergizes with ionizing radiation to inhibit tumor growth(A) PSN1 xenografts (PSN1 cells co-injected with LTC-14 stellate cells) established in the flank of athymic nude mice were treated with MK-2206 (60 mg/kg-320 mg/kg) as indicated or beta -cyclo-dextrin (1.5 mg/ml) carrier. Xenograft tumors were lysed & lysates probed by western blot with the indicated antibodies. (B-D) Sections of PSN1 xenografts treated with three consecutive doses of MK-2206 (60 mg/kg). (B) Sections of PSN1 xenografts & in-vitro PSN1 cells fixed & stained with anti-NPM (red) & anti-p14ARF (green). (C) PSN1 xenografts treated with MK-2206 or carrier were stained with DAPI, anti-p53 (DO1) or p53mut (OP29 clone) (D) PSN1 xenografts treated with MK-2206 (60 mg/kg) or carrier were stained by immunohistochemical methods with anti-pS473-AKT, anti-phospho-S48-NPM (pS48-NPM) or p53. (E) PSN1 xenografts established in the flank of athymic nude mice were injected subcutaneously with two alternate day doses of MK-2206 (60 mg/kg) or carrier. Mice were subsequently treated with a single dose of IR (6 Gy) & tumor volumes measured regularly with callipers. Dash lines indicate tumor growth differential at 250 mm3. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p14ARF/CDKN2A Antibody - BSA Free [NB200-111] - Inhibition of AKT promotes enhanced MDM2 activity via the increased association between NPM & p14ARF(A) Npm−/−, p53−/−double null MEF were infected with pBABE retrovirus empty vector & pBABE expressing FLAG-tagged-NPM-WT, NPM-S48A or S48E as indicated. Immunopurification of NPM was done by pulling down with the Flag tag (middle panel) followed by elution of complexes by the Flag peptide & subsequent immunopurification of endogenous MDM2 (lower panel). (B) Nuclear immunoprecipitates of MDM2 from T24 cells treated with MK-2206 (5 μM, 24 hrs). Immunoprecipitates & lysates were blotted with the indicated antibodies. (C) T24 cells were treated with MK-2206 (5 μM) as indicated. p14ARF was immunoprecipitated from whole cell lysates & nuclear extracts & the association with NPM & MDM2 determined by western blot. Immunoprecipitates & lysates were blotted with the indicated antibodies. (D) MDM2 & (E) p53 ubiquinitation assay in H1299 cells transfected with wild type p53, HA-tagged ubiquitin & treated for 16 hrs with DMSO, MK-2206 (5 μM) or Nutlin3A (5 μM) as indicated. Immunoprecipitates & whole cell lysates were probed with the indicated antibodies. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25071014), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

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	NB200-111AF350
Alexa Fluor 405	NB200-111AF405
Alexa Fluor 488	NB200-111AF488
Alexa Fluor 532	NB200-111AF532
Alexa Fluor 594	NB200-111AF594
Alexa Fluor 647	NB200-111AF647
Alexa Fluor 700	NB200-111AF700
Alexa Fluor 750	NB200-111AF750
Allophycocyanin	NB200-111APC
Biotin	NB200-111B
CoraFluor 1	NB200-111CL1
DyLight 350	NB200-111UV
DyLight 405	NB200-111V
DyLight 488	NB200-111G
DyLight 550	NB200-111R
DyLight 594	NB200-111DL594
DyLight 650	NB200-111C
DyLight 680	NB200-111FR
DyLight 755	NB200-111IR
FITC	NB200-111F
HRP	NB200-111H
Janelia Fluor 525	NB200-111JF525
Janelia Fluor 549	NB200-111JF549
Janelia Fluor 646	NB200-111JF646
Janelia Fluor 669	NB200-111JF669
PE	NB200-111PE
PerCP	NB200-111PCP
mFluor Violet 450 SE	NB200-111MFV450
mFluor Violet 500 SE	NB200-111MFV500
mFluor Violet 610 SE	NB200-111MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB200-111SS

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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

Marker

Modification

Clonality

Clone

Gene Symbol

Images Type

Isotype

Kit Type

Lysate Type

Preservative

Protein Or Peptide

Research Areas

Has Sample Size

Promotional Item

Species Glossary

LC3B Antibody - BSA Free

LC3B Antibody - BSA Free

Stathmin-2/STMN2 Antibody - BSA Free

GADD153/CHOP Antibody (9C8) - BSA Free

Ubiquitin Antibody (Ubi-1) - BSA Free

beta-III Tubulin Antibody - BSA Free

SLC31A1/CTR1 Antibody - BSA Free

GADD153/CHOP Antibody - BSA Free

NF-H Antibody

USP7 Antibody - BSA Free

Mas Antibody - BSA Free

p14ARF/CDKN2A Antibody - BSA Free