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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
Alexa Fluor Plus 405	NB600-1384AFP405
Alexa Fluor Plus 488	NB600-1384AFP488
Alexa Fluor Plus 555	NB600-1384AFP555
Alexa Fluor Plus 594	NB600-1384AFP594
Alexa Fluor Plus 647	NB600-1384AFP647
Alexa Fluor Plus 680	NB600-1384AFP680
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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293-VE

Recombinant Human VEGF 165 Protein

(469 Publications)

10 μg

293-VE

As an alternative, please consider our next generation Recombinant Human VEGF 165 (Catalog # BT-VEGF). It has equivalent bioactivity to Recombinant Human VEGF 165 (Catalog # 293-VE). It combines R&D Systems quality with scalability that allows for a solid supply chain. Both Recombinant Human VEGF 165 proteins are measured in a cell proliferation assay using HUVEChuman umbilical vein endothelial cells.

1 μg/lane of Recombinant Human VEGF165was resolved by SDS-PAGE with silver staining, under reducing (R) and non-reducing (NR) conditions, showing major bands at 20-22 kDa and 39-42 kDa, respectively. Multiple bands in gel are due to glycosylation.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	293-VE-001MG/CF
Carrier Free	293-VE-010/CF
Carrier Free	293-VE-050/CF
Carrier Free	293-VE-500/CF

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NB300-221

GAPDH Antibody (1D4) - BSA Free

(19 Reviews)

(455 Publications)

0.1 ml

NB300-221

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

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

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

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

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

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

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

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

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

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

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

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

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

Reactivity:

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

Species Glossary

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

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB300-221SS

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D8000C

Human IL-8/CXCL8 Quantikine ELISA Kit

(463 Publications)

1 Kit

D8000C

RT-qPCR for a set of cytokines (CSF2, IL6, IL8 and IL1B) and chemokines (CXCL1 and CXCL6) of total RNA samples derived from mono- and co-cultures in a transwell assay.The respective co- (+) or mono- (-) culture is indicated on the X-axis. The respective analyzed cytokine or chemokine is indicated in the header of each graph. Expression values are shown in arbitrary units (AU) and have been normalized to beta-2 microglobulin (B2M) mRNA copies. Experiments were performed in triplicates (n = 3). Statistical analysis was performed on the mean values by unpaired comparison of mono-cultured HDF and co-cultured HDF RNA samples by using Student’s t-test (**p<0.01, ***p<0.001; n.s.: not significant). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25919140), licensed under a CC-BY license. Not internally tested by R&D Systems.

Pathogen-associated molecular patterns (PAMPs) and damage-associated molecular pattern (DAMPs) induce cytokine expression in human breast cancer cells in vitro. a Relative expression of IL-6 and IL-8 mRNAs after 6 h stimulation with lipopolysaccharide 1 (LPS1) and LPS2, using quantitative real-time PCR (QPCR) on mRNA from the cell lines indicated; n = 6. Error bars standard error of the mean (SEM); ***P <0.001 (analysis of variance (ANOVA)). b Release of IL-6 and IL-8 after stimulation with LPS1 and cycloheximide for 6 h, using ELISA on supernatants from stimulated MDA-MB-231 cells; n = 3. c-e Relative release of IL-6 and IL-8 after stimulation with LPS1, LPS2, the DAMP HMGB1 or IL-1 beta for 6 h, using ELISA on supernatants from stimulated MDA-MB-231 cells (c), SUM-149 cells (d) and SUM-159 cells (e); n = 10. Error bars SEM; *P <0.05, ***P <0.001 (ANOVA). f Dual luciferase reporter assays of MDA-MB-231 cells transfected with an NF kappa B reporter. TK-Renilla was co-transfected as control (Ctrl). LPS1, LPS2, HMGB1 or IL-1 beta was added to stimulate NF kappa B activity as described in “Methods”; n = 14. Error bars SEM; ***P <0.001 (ANOVA). g The relative release of IL-6 and IL-8 after stimulation with the DAMP S100A9 for 20 h, using ELISA on supernatants from stimulated MDA-MB-231, SUM-149 and SUM-159 cells; n = 6. Error bars SEM. *P <0.05, ***P <0.001 (ANOVA). h Transient transfection of pDUO-MD2/hTLR4 for 72 h in MCF-7 cells induces release of both IL-6 and IL-8 measured by ELISA; n = 6. Error bars SEM; *P <0.05, ***P <0.001 (Students t test) Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26392082), licensed under a CC-BY license. Not internally tested by R&D Systems.

Toll-like receptor 4 (TLR4) silencing decreases endogenous levels of pro-inflammatory cytokines. a Effect of TLR2/4 silencing in breast cancer cells transfected with negative control (nc) siRNA, or siRNA directed against TLR2 mRNA (si#1 and si#2) or TLR4 mRNA (si#1 and si#2) was analyzed using quantitative real-time PCR; ***P <0.001 (analysis of variance (ANOVA)). b IL-6 (left) and IL-8 (right) ELISA on supernatants from MDA-MB-231 breast cancer cells transfected with nc siRNA, or siRNA directed against TLR2 mRNA (si#1 and si#2) or TLR4 mRNA (si#1 and si#2); n = 4. Error bars standard error of the mean (SEM); *P <0.05, **P <0.01, ***P <0.001 (ANOVA). c Boyden chamber migration assays. Migration of primary human myeloid cells towards supernatants from different cell lines indicated. Human primary peripheral blood mononuclear cells were isolated as previously described [48] and allowed to migrate through a Costar Transwell® Permeable Support 8.0-μm 24-well plate (Corning) to the supernatants of breast cancer supernatants cultured under serum-free conditions. Percentage of migrated CD11b+ cells was analyzed using a flow cytometer and CD11b-APC antibodies (BD Sciences); n = 4. Error bars SEM; *P <0.05, **P <0.01, ***P <0.001 (ANOVA). d Matrigel invasion assays. Invasion of lipopolysaccharide (LPS)-stimulated/un-stimulated MDA-MB-231 cells into matrigel invasion chambers (BD Sciences) as indicated: 25 × 103 MDA-MB-231 cells were stimulated or not with LPS and allowed to invade from 72 h. Amount of invaded cells was analyzed using crystal violet staining and manual counting in four separate experiments; n = 4. Error bars SEM; *P <0.05, **P <0.01, ***P <0.001 (Student’s t test) Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26392082), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Conjugate	Catalog #	Availability	Size	Price
HRP	PD8000C
HRP	S8000C

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AF2018

SOX2 Antibody [Unconjugated]

(6 Reviews)

(454 Publications)

100 μg

AF2018

Western blot shows lysates of D3 mouse embryonic stem cell line, NTera-2 human testicular embryonic carcinoma cell line, F9 mouse teratocarcinoma stem cells, and rat cortical stem cells. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for SOX2 at approximately 36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

BG01V human embryonic stem cells were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. SOX2/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) or control antibody (AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (MAG999) and DNA was purified using chelating resin solution. Thenanogpromoter was detected by standard PCR.

SOX2 was detected in immersion fixed undifferentiated mouse cortical stem cells using Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; (NL001) and counterstained with DAPI (blue). Nestin was also detected in stem cells using Mouse Anti-Mouse/Rat Nestin Monoclonal Antibody (MAB2736) and co-stained using the NorthernLights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; NL009). Specific staining of SOX2 was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

SOX2 was detected in immersion fixed undifferentiated rat cortical stem cells using Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Nestin was also detected using Mouse Anti-Mouse/Rat Nestin Monoclonal Antibody (MAB2736) and stained using the NorthernLights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; NL009). Specific staining of SOX2 was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Simple Western lane view shows lysates of BG01V human embryonic stem cells and NTera-2 human testicular embryonic carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for SOX2 at approximately 53 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

SOX2 was detected in immersion fixed ADLF1 (top panel) and FAB2 (bottom panel) induced pluripotent stem cell lines using Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

SOX2 was detected in immersion fixed paraffin-embedded sections of glioblastoma using Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei in cancer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

SOX2 was detected in immersion fixed NTera‑2 human testicular embryonic carcinoma cells (Positive) & absent in HepG2 human hepatocellular carcinoma cells (Negative) using Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to Cell surface and nuclear. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

<b>Genetic Strategies Validation. </b>Knockdown OGT inhibits proliferation and tumorsphere formation of hepatoma cell through reducing eIF4E expression. A, Huh7 and PLC/PRF/5 cells were infected with control shRNA, OGT shRNA2 alone, or with wild‐type eIF4E lentivirus. The cell lysates were harvested for western blotting analysis using indicated antibodies. beta ‐actin expression was served as a loading control. B, Cell proliferation of Huh7 and PLC/PRF/5 cells infected with lentiviruses as in panel (A) were measured with CCK8 assay. (C‐H) Huh7 and PLC/PRF/5 cells infected with lentiviruses as in panel (A) were seeded into 96‐well plates. After 12 d, tumorsphere were counted and quantified. Representative images of sphere (scale bars, 100 μm) were shown (C, F). The diameter of sphere (D, G) and number of sphere (E, H) were count. Data represent mean ± SD of at least three independent experiments. The two‐tailed Student's t tests were used. **P < 0.01. I, Huh7 cells expressing either OGT shRNA2 alone or with wild‐type eIF4E lentivirus were incubated with PE‐labelled anti‐AC133 antibody. The percentages of CD133+ cells in graphs were analysed by flow cytometry. Black line, control IgG staining; red line, CD133 staining. J, Cell lysates were examined by western blotting with indicated antibodies. The right panel showcases relative protein amounts of different groups. Error bars represent ±SD of triplicate experiments. **P < 0.01; n.s, no significance. K, Huh7 cells were collected and subjected to immunoprecipitation with antibody against eIF4E or normal mouse IgG. Total RNAs were purified from immunocomplexes and subjected to RT‐PCR to measure Sox2, OCT4, and KLF4 mRNAs associated with eIF4E Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30677218), licensed under a CC-BY license. Not internally tested by R&D Systems.

TP63 and SOX2 staining in cervical biopsies.Immunohistochemical staining of cervical biopsies. Bars = 50µm.(A) normal cervix no primary control; (B) normal cervix stained with anti-SOX2; (C) Squamous cell cervical carcinoma no primary control; (D) representative TP63, and (E) representative SOX2, staining of tumour cells. For both TP63 and SOX2 staining was seen in the nucleus of positive cells (examples indicated by solid arrows); negative cells were a minority of tumour cells (examples indicated by unfilled arrows). (F) Image analysis results of % nuclear +ve tumour cells in biopsies. Parallel sections from 11 cases were stained with SOX2 and TP63. Tumour cells were evaluated for their nuclear expression of the transcription factors. There was no significant difference between the data for SOX2 and TP63 (Wilcoxon signed rank test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25531390), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Biological Strategies Validation. </b>Direct effects of glucocorticoids in ependymal glia. (A–I) Zebrafish spinal cord transverse sections. (A–C) Immunostaining of wild-types for Nr3c1 (green) and Gfap (red), plus DAPI staining (blue) with Dex treatment at 24 h post sham injury (A), 24 h (B) and 48 h post SCI (C). Nuclear Nr3c1 (active) localization in Gfap-positive cells around the central canal is stimulated with Dex treatment. (D–F) Immunostaining for EGFP (pan-cytoplasmic) in the SR4G transgenic reporter line (green) and filamentous Gfap (red) at 24 h post sham injury (D) and SCI with no treatment (E) or 24 h post SCI +Dex (F). Nr3c1 signaling is constitutively active in ependymal glia in uninjured controls (D), diminished after SCI (E), and was maintained after SCI + Dex. (G–I) Immunostaining for Sox2 (green), PCNA (red) and Gfap (blue) from 48 h post sham injury (G), 48 h post SCI followed by no treatment (H), or 48 h post SCI +Dex (I). Sox2 immunoreactivity is reduced in ependymal glia and neural precursor cells after Dex treatment. Representative images from 10 spinal cords per time point in each condition. Scale bars, 20 μm (A is the same as B,C; D is the same as E,F; G is the same as H,I). ∗denotes central canal. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31069223), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Biological Strategies Validation. </b>Epigenetic perturbation of enhancer clustering and genome-widebinding.(A) The fluctuation range (x) and amplitude (y) were obtainedby fitting the pair-correlation function of the indicated dataset with thefluctuation model. Figure 2 and Figure 2—figure supplement 1,Equations10–13. Supplementary file 1. Data from the same conditionwere grouped in separate ellipses. (B) Sox2 ChIP-exo peakdensity distribution in the wild-type and TSA treated (red dotted) cellsacross chromosome 1, 2, 3. In the upper panels, each chromosome was dividedto 500 bins. The color map correlates with the number of peaks in each bin.Top 7000 binding sites were considered in each condition. (C)Cumulative density histogram of the distances to transcription start sites(TSS's) of Sox2 ChIP-exo peaks in WT, Sox2 ChIP-exo peaks in the TSA treatedcells (red dotted), and random genomic positions (gray).DOI://dx.doi.org/10.7554/eLife.04236.013 Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25537195), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Genetic Strategies Validation. </b>Generation of KCNA5 knockout model. (A) Illustration of polycistronic self-replicating RNA used for reprogramming (Yoshioka et al., 2013). Reading frames for OCT4, KLF4, SOX2, GLIS1, and puromycin resistance are separated by 2A peptide-encoding or IRES sequences. (B) Phase contrast morphology (5 x) and alkaline phosphatase staining of the F1 wild-type hiPSC line generated using the RNA vector shown in (A). (C) Representative F1 karyogram indicating a normal male genotype. (D) The RNA vector is undetectable in established F1 hiPSCs following puromycin withdrawal, whereas freshly transfected parental fibroblasts display robust transgene expression (RT-qPCR data, n = 2). (E) Bisulfite sequencing of OCT4 and NANOG promoter regions of the indicated samples showing an hESC-like hypomethylated state in F1 cells. (F) RT-qPCR analysis of F1 hiPSCs in comparison to the original fibroblasts and a hESC control (n = 2). (G) Immunostains showing robust pluripotency factor expression in F1 hiPSCs at the protein level. (H) Illustration of the Kv1.5 channel. The voltage sensor in transmembrane helix S4 is indicated by symbols. The ion pore region is in-between segments S5 and S6. (I)KCNA5 knockout strategy based on simultaneously transfecting 4 CRISPR/Cas9 nickase vectors targeting the 5' end of the 1-exon gene body. A mutant clone generated this way displayed homozygous deletions causing a frame shift and premature stop as illustrated at the bottom. (J) PCR spanning the mutation site validates the sequencing-verified frame shift-causing deletion in KCNA5fs/fs cells at the DNA and RNA levels. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fphys.2017.00469/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Construction and expression of srRNA.A: Scheme of Self-Replicating srRNA constructs. B: Expression of reprogramming factors in OKS-iM 4F-srRNA, OKS-iG 4F-srRNA, OKS-iGM 5F-srRNA, and OKS-iGML 6F-srRNA on day 2 (left) and day 10 (right). BJ cells were co-transfected with srRNAs plus B18R mRNA (1:1 srRNA:B18R mRNA), selected with puromycin and cultured in Advanced DMEM/10% FBS for 10 days. C: qRT-PCR analysis of LIN28A and NANOG from 4F-srRNA and 5F-srRNA transfected BJ cells normalized to GAPDH. Cells were selected with puromycin after the transfection, and cultured in Advanced DMEM/10% FBS (Ad) or ES medium. Cells were collected on day 2, 4, 6, 8 and 10 for 5F-srRNA, and day 10 for 4F-srRNA (OKS-iM and OKS-iG). D: Immunoblot analysis of LIN28A and NANOG in 5F-srRNA transfected BJ cells on day 6 and 10. iPSC: iPSC clone generated with 5F-srRNA from BJ cells. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28750082), licensed under a CC-BY license. Not internally tested by R&D Systems.

The Majority of Ferret and Human, but Not Mouse, Sox2-Positive Tbr2-Negative BPs Exhibit Nuclear YAP. (G and H) that are shown at higher magnification (F′, F″, G′, G″, H′, and H″), as indicated; selected Tbr2-positive nuclei that are YAP negative in mouse, ferret, and human are outlined by white lines.(I–K) Quantification of the percentage of Tbr2-negative nuclei in the VZ (I), Tbr2-positive nuclei in the VZ (J), and Tbr2-positive nuclei in the SVZ (K) that are YAP positive in mouse E14.5, ferret E36, and human 11 wpc neocortex. Two or three images per embryo-fetus were taken, 30 randomly picked Tbr2-negative nuclei in the VZ (I) and Tbr2-positive nuclei in the VZ (J) and SVZ (K) were scored per image, and the values obtained were averaged for each embryo-fetus. Data are the mean of three or four embryos-fetuses.(A–C and F–H) Images are 1-μm optical sections. Scale bars represent 50 μm (A–C and F–H), 10 μm (A′, B′, and C′), and 20 μm (F′, F″, G′, G″, H′, and H″).(D, E, and I–K) Error bars indicate SEM; ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA, post hoc Tukey HSD).See also Figures S1 and S2. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31018127), licensed under a CC-BY license. Not internally tested by R&D Systems.

The Majority of Ferret and Human, but Not Mouse, Sox2-Positive Tbr2-Negative BPs Exhibit Nuclear YAP(A–C) Double immunofluorescence for YAP (green) and Sox2 (magenta), combined with DAPI staining (white), of mouse E14.5 (A), ferret E36 (B), and human 14 wpc (C) neocortex. Boxes indicate areas in the SVZ (A) and oSVZ (B and C) that are shown at higher magnification (A′, B′, and C′); selected Sox2-positive nuclei that are YAP negative in mouse and YAP positive in ferret and human are outlined by white lines; arrowheads indicate a YAP-positive basal process of a bRG. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31018127), licensed under a CC-BY license. Not internally tested by R&D Systems.

The Majority of Ferret and Human, but Not Mouse, Sox2-Positive Tbr2-Negative BPs Exhibit Nuclear YAP .(F–H) Double immunofluorescence for YAP (green) and Tbr2 (magenta), combined with DAPI staining (white), of mouse E14.5 (F), ferret E36 (G), and human 11 wpc (H) neocortex. Boxes indicate areas in the VZ and SVZ (F) or iSVZ (G and H) that are shown at higher magnification (F′, F″, G′, G″, H′, and H″), as indicated; selected Tbr2-positive nuclei that are YAP negative in mouse, ferret, and human are outlined by white lines.(I–K) Quantification of the percentage of Tbr2-negative nuclei in the VZ (I), Tbr2-positive nuclei in the VZ (J), and Tbr2-positive nuclei in the SVZ (K) that are YAP positive in mouse E14.5, ferret E36, and human 11 wpc neocortex. Two or three images per embryo-fetus were taken, 30 randomly picked Tbr2-negative nuclei in the VZ (I) and Tbr2-positive nuclei in the VZ (J) and SVZ (K) were scored per image, and the values obtained were averaged for each embryo-fetus. Data are the mean of three or four embryos-fetuses.(A–C and F–H) Images are 1-μm optical sections. Scale bars represent 50 μm (A–C and F–H), 10 μm (A′, B′, and C′), and 20 μm (F′, F″, G′, G″, H′, and H″).(D, E, and I–K) Error bars indicate SEM; ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA, post hoc Tukey HSD).See also Figures S1 and S2. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31018127), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunohistochemical analysis of Trp53-/-Notch1-/-Notch2-/- and Trp53-/-Rbpj-/- tumors. (A) A Trp53-/-Notch1-/-Notch2-/- tumor in the SVZ. Streams of tumor cells invading the brain parenchyma are indicated by arrowheads. The GFP expression from the Rosa-CAG::GFP Cre-reporter indicates derivation from Hes5+ cells. (B) Expression of GFP, progenitor (SOX2) and glial markers (OLIG2, GFAP), as well as staining for mitotically active cells (PCNA) and immature neurons (DCX) in Trp53-/-Notch1-/-Notch2-/- and Trp53-/-Rbpj-/- tumors. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36358826), licensed under a CC-BY license. Not internally tested by R&D Systems.

Deregulation of SOX2, ASCL1&OLIG2 TFs in p27KO cultures at the onset of differentiation. e Immunocytochemistry for p27 (red) in combination with ASCL1, OLIG2 or SOX2 (green) in 2 + 1DIV NPCs. Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36627412), licensed under a CC-BY license. Not internally tested by R&D Systems.

Padi2/3 DKO TSCs exhibit diminished stem cell potential&are prone to differentiate into large trophoblast giant cells (TGCs). (D) Immunofluorescence staining on WT&Padi2/3 DKO TSCs for trophoblast stem cell markers CDX2&SOX2. Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36010543), licensed under a CC-BY license. Not internally tested by R&D Systems.

Unedited images from samples featured in Figure 6.Stage 39–40 larvae were amputated and processed for whole-mount immunostaining at the indicated hour post amputation (hpa), with the exception of the 0 h group, which was first fixed and then amputated to represent the pre-amputation group. Images show representative total z-projections of whole-mount immunostained samples for each group corresponding to the edited images shown in Figure 6. Transverse red dashed line indicates amputation plane. Scale bar, 100 µm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33955353), licensed under a CC-BY license. Not internally tested by R&D Systems.

Notch1 and Trp53 deletion in Hes5+ cells leads to formation of forebrain tumors in adult mice. (A) Kaplan–Meier curves showing survival of Hes5::CreERT2 Trp53-/-Notch1-/- and Trp53-/-Notch2-/- mutant mice. (B) A Trp53-/-Notch1-/- tumor in the forebrain. The GFP expression from the Rosa-CAG::GFP Cre-reporter indicates derivation from Hes5+ cells. (C) Percentages of Trp53-/-Notch1-/- and Trp53-/-Notch2-/- mice that developed hyperplasia or tumors in the brain. (D) A Trp53-/-Notch1-/- tumor (dashed line) in the anterior forebrain. In contrast, the forebrain of Trp53-/-Notch2-/- mice appears grossly normal. (E) Expression of GFP, progenitor (SOX2) and glial markers (OLIG2, GFAP), as well as staining for mitotically active cells (PCNA) and immature neurons (DCX) in Trp53-/-Notch1-/- tumors. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36358826), licensed under a CC-BY license. Not internally tested by R&D Systems.

Wnt signaling downregulates Tcf3 expression in mouse ESCs.A. qRT-PCR analysis of Tcf3 in wild type, ApcNN and ApcMin/Min ESCs. Actb was used as an internal control; bars represent n = 2 ± SD. B. Western blot analysis of the core pluripotency markers Oct4, Nanog, Sox2 and Tcf3 on protein lysates isolated from two independent ApcNN clones and wild type control ESCs. Actb and Tubulin were used as an internal control. C–D. qRT-PCR analysis of Tcf3 in wild type ESCs treated for different time intervals with Wnt3a conditioned medium (C), or with the GSK-inhibitor SB-216763 (D). L-medium and DMSO were employed as controls, respectively. Actb was used as an internal control; bars represent n = 2 ± SD. E. Time course western blot analysis of Tcf3 expression in wild type ESCs treated with Wnt3a conditioned medium (Wnt3a CM) or control L-medium (LM). Actb was used as an internal control. F. qRT-PCR analysis of Tcf3 and Wnt target genes Axin2 and Cdx1 in wild type ESC treated for 48 h with different concentrations of GSK-inhibitor SB-216763 or DMSO as control. Actb was used as an internal control; bars represent n = 2 ± SD. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23658527), licensed under a CC-BY license. Not internally tested by R&D Systems.

Fetal rat lung stained for Sox2 (red), co-stained with DAPI (blue). Image from a verified customer review.

SOX2, OCT2, and PRRX1 are downstream effectors of the SETD2‐regulated EMT program. (A) Expression of SOX2, OCT2, and PRRX1 in TGF‐ beta ‐treated WT (72 h), SETD2 KO, and SETD2 rescue tested by RT‐qPCR (run in triplicate). (B) Migration capacity by wound healing assay, (C) invasiveness by transwell assay, and (D) stemness by 3D spheroid formation assay in RPTEC WT GFP (control vector), SETD2 KO1 and KO2, and SOX2/OCT2/PRRX1‐transduced WT RPTEC lines. Images are taken at 4× magnification, scale bar: 1000 μm for (B) and (D) and at 2.5× magnification, scale bar: 1200 μm for (C). Data are represented as mean ± SEM for triplicate reactions for B–D. P‐value is calculated by one‐way ANOVA in (A), (B), and (D). Two‐way ANOVA is used for statistical test for (C). ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05; ns, P ≥ 0.05. (E) Model of the SETD2 loss‐driven EMT program through cell intrinsic (transcriptional) and cell extrinsic (paracrine) mechanisms. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/37418588), licensed under a CC-BY license. Not internally tested by R&D Systems.

Temporally coordinated expression of Sox2, Tbr2, NueroD, and Prox1 in gfap-GFP+ GNPs and differentiating granule neurons Gfap-GFP+/Sox2+ progenitor cells were found at the dentate neuroepithelium at E14 (arrow in A1). Sox2 expression was found at the VZ. Tbr2+ GNPs are then observed (red, arrow in A2). Tbr2 expression is followed by NeuroD expression (A3). Tbr2+/NeuroD+ GNPs are found (arrow, A3). The Tbr2+ GNPs (red) migrate toward the DG primordium and co-express Prox1 (green), a marker for early differentiating dentate granule neurons (arrow in A4). Gfap-GFP+ progenitors sequentially express Sox2, Tbr2, NeuroD1, and then, Prox1 in the developing DG at E16 (arrows in B1–B4, respectively). Some Sox2+/Tbr2+, Tbr2+/NeuroD+, and Tbr2+/Prox1+ GNPs are found. Similarly, gfap-GFP+ progenitor cells contribute to Tbr2+/NeuroD+ (arrows in D1,D2), Prox1+ GNPs (arrows in D3,D4) at P3. Schematic drawing of the expression patterns of gfap-GFP and transcription factors during the differentiation of GNPs (E). Scale bars; 200 μm in (A1–A4, B1–B4, C1–C3, D1,D3); 50 μm in (D2,D4). Image collected and cropped by CiteAb from the following open publication (//www.frontiersin.org/articles/10.3389/fnins.2024.1425849/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , ,

Applications: , , , ,

Host:

Goat Polyclonal

Host: Goat 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	AF2018U-100UG
Alexa Fluor 405	AF2018V-100UG
Alexa Fluor 488	AF2018G-100UG
Alexa Fluor 532	AF2018X-100UG
Alexa Fluor 594	AF2018T-100UG
Alexa Fluor 647	AF2018R-100UG
Alexa Fluor 700	AF2018N-100UG
Alexa Fluor 750	AF2018S-100UG
Allophycocyanin	AF2018A
Biotin	BAF2018
CoraFluor 1	AF2018CL1
DyLight 350	AF2018D
DyLight 405	AF2018E
DyLight 488	AF2018K
DyLight 550	AF2018L
DyLight 594	AF2018M
DyLight 650	AF2018W
DyLight 680	AF2018Y
DyLight 755	AF2018Z
FITC	AF2018F
HRP	AF2018H
Janelia Fluor 525	AF2018JF525
Janelia Fluor 549	AF2018I
Janelia Fluor 646	AF2018J
Janelia Fluor 669	AF2018JF669
NL493	NL20181G
PE	AF2018P
PerCP	AF2018C
mFluor Violet 450 SE	AF2018MFV450
mFluor Violet 500 SE	AF2018MFV500
mFluor Violet 610 SE	AF2018MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	NL20181G
Carrier Free	AF2018-SP

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7270-IL

Recombinant Human IL-6 (HEK293-expressed) Protein

(10 Publications)

25 μg

7270-IL

Measured in a cell proliferation assay using T1165.85.2.1 mouse plasmacytoma cells. The ED50 for this effect is 0.1-0.6 ng/mL.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	7270-IL-025/CF
Carrier Free	7270-IL-010/CF
Carrier Free	7270-IL-100/CF

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DB100C

Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA

(453 Publications)

1 Kit

DB100C

Reactivity:

Hu, Mu, Rt, Po, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca

Applications:

Applications:

Conjugate	Catalog #	Availability	Size	Price
HRP	SB100C
HRP	PDB100C

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	SB100C
Carrier Free	PDB100C

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H00008878-M01

p62/SQSTM1 Antibody (2C11) - Azide and BSA Free

(9 Reviews)

(441 Publications)

0.1 mg

H00008878-M01

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - ICC-IF analysis of SQSTM1 protein in HeLa cell using SQSTM1/p62 antibody (clone 2C11) at 10 ug/ml concentration.

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Analysis of p62 in SH-SY5Y cells using anti-p62/SQSTM1 antibody. Red - p62 puncta; Blue - nuclear DAPI; Green - Cytoskeleton. Image from verified customer review.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Disruption of the VANGL2-p62/SQSTM1 interaction in breast cancer cells. Soft agar colony formation of T47D cells overexpressing GFP, GFP-VANGL2 and GFP-p62/SQSTM1 (right). Protein expression was revealed with anti-p62/SQSTM1, anti-VANGL2 and anti-GFP antibodies by western blot analysis (right). In anti-GFP blot, the arrowhead indicates position of co-migrating GFP-VANGL2 and GFP-p62/SQSTM1 and the asterisk pinpoints GFP alone. Error bars represent mean+/-s.d. (n=3). Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms10318), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Analysis of p62/SQSTM1 antibody on HeLa cells. Image from verified customer review.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Confirmation of loss of hepatic ATG7 protein and a phenotype of hepatic autophagy inactivation. Fourth ug of liver protein were loaded on 4-20% gradient Tris-Glycine gels and subsequently immunoblotted with antibodies against ATG7, p62 and LC3. Blots were visualized with G:BOX Chemi XRQ (SynGene). Representative immunoblots are shown for WT and KO mice. Full-length immunoblots are presented in Supplementary Figure S1. ATG7: autophagy-related protein 7; p62: sequestosome 1; LC3: microtubule-associated protein 1 light chain 3; WT: Mx1-Cre-Atg7F/F mouse; KO: Mx1-Cre+Atg7F/F mouse. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-14405-w), licensed under a CC-BY license.

ELISA: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Detection limit for recombinant GST tagged SQSTM1 is approximately 0.03ng/ml as a capture antibody.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Activation of multiple signaling pathways by dandelion root extractA. Western blots of proteins involved in programmed cell death & cell survival & inflammation B. Densitometry quantification of western blot analysis from three independent experiments. *P<0.05, **P< 0.001. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.11485), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Elevated levels of additional proteins & GM3 ganglioside in the MEC of MPS IIIB brain.Staining performed with antibodies to the indicated substances was observed in the MEC region of 3 month-old MPS IIIB mice (for total ubiquitin & polyubiquitin) & 6 months for all others. Staining was not seen in the MEC region of age-matched control mice (Naglu +/−) nor in the LEC region of MPS IIIB mice (the latter not shown). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0027461), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Confirmation of loss of hepatic ATG7 protein & a phenotype of hepatic autophagy inactivation. Fourty µg of liver protein were loaded on 4–20% gradient Tris-Glycine gels & subsequently immunoblotted with antibodies against ATG7, p62 & LC3. Blots were visualised with G:BOX Chemi XRQ (SynGene). Representative immunoblots are shown for WT & KO mice. Full-length immunoblots are presented in Supplementary Figure S1. ATG7: autophagy-related protein 7; p62: sequestosome 1; LC3: microtubule-associated protein 1 light chain 3; WT: Mx1-Cre−Atg7F/F mouse; KO: Mx1-Cre+Atg7F/F mouse. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29074879), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - The content of mitochondrial p62, OPTN, & VCP in spinal cord of SOD1‐G93A mice is decreased by Parkin knockoutImmunostaining of anterior horn of lumbar spinal cord for ChAT (in green) & p62 (in red) at 130 days. Scale bar, 10 μm.Quantification of the percentage of ChAT‐positive motor neurons (MN) containing p62‐positive inclusions at 130 days. PKO/G93A MN have less SOD1 aggregates than G93A, while Non Tg & PKO MN have none. Data were collected from n = 4 (two males & two females) mice for Non Tg & PKO & n = 6 (three males & three females) for G93A & PKO/G93A. Number of images recorded for each genotype: n = 9 for Non Tg & PKO, n = 16 for G93A, & n = 12 for PKO/G93A. Results are expressed as mean ± SEM; *P = 0.032, by unpaired one‐tailed Student's t‐test.Representative Western blots of OPTN, p62, & VCP in spinal cord mitochondrial fractions at 130 days. Protein levels are normalized by Complex V. The quantifications in panels (D–F) show that mitochondria of PKO/G93A mice have less mitophagy adaptor proteins than G93A relative to Complex V.Quantification of OPTN relative to Complex V at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); *P = 0.039 by paired Student's t‐test.Quantification of p62, relative to Complex V at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); **P = 0.0029 by paired Student's t‐test.VCP was quantified using Complex V as loading reference at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); *P = 0.044 by paired Student's t‐test.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30126943), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - miR-29a downregulates TFEB & ATG9A to inhibit autophagyA. Schematic representation of the miR-29 family members & 3′-UTR binding sites of miR-29 targets as well as mutated binding sites used in Luciferase Assays: Transcription Factor EB (TFEB) & Autophagy-related protein 9A (ATG9A). All three miR-29 family members (miR-29a, miR-29b, & miR-29c) have identical seed sequences. Conserved miR-29 binding sites in the 3′-UTR of mRNA transcripts encoding ATG9A & TFEB are depicted in bold. B. 10ug of total protein cell lysates from Panc-1 transfected with CTRL or miR-29a mimics were subjected to western blot analysis for TFEB, ATG9A, & GAPDH. Relative quantification of band intensities normalized to GAPDH are shown below respective blots. C. Relative firefly luciferase activity from TFEB & ATG9A 3′ UTR wild type (WT) & mutant (mut) reporter constructs following co-transfection into Panc-1 cells with control or miR-29a mimics. All readouts were normalized to renilla luciferase activity for each well. Average relative luminesce normalized to respective controls is presented (n=6) ± S.E.M. D. 5ug of total protein cell lysates from Panc-1 cells were transfected with CTRL, miR-29a mimics, siCTRL, siTFEB, or siATG9A. 24 hours post-transfection, total protein was harvested & subjected to western blot analysis for p62 & LC3B, & GAPDH was used as loading control. Quantification of band intensities normalized to GAPDH & relative to control are shown below respective blots. All experiments were repeated 3 times & representative data is presented. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.11928), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Effects of autophagy inhibitors in combination with IFN-gamma on expression of PD-L1 in gastric cancer cell lines. a The effect of chloroquine (CQ) or 3-MA on expression of PD-L1 with or without INF-gamma for 24 h was determined by flow cytometry assays. In AGS & NCI-n87 cells, MFI as the indication of PD-L1 expression level can be further increased by the treatment of INF-gamma . b Levels of LC3B-I/II, p62/SQSTM1 & PD-L1 were determined by Western blots in AGS & NCI-n87 cells treated by CQ, 3-MA, bafilomycin A1 (Baf) or rapamycin (Rap) for 24 h. c Positive staining of PD-L1 (red) & LC3 positive puncta (green) was determined by immunofluorescence in AGS & NCI-n87 cells treated by autophagy inhibitors & activator as in (b). d Rapamycin decreased the levels of PD-L1 protein in AGS & NCI-n87 cells as shown by flow cytometry. Results were averaged & blots were representative of 4 independent experiments, *p < 0.05, **p < 0.01 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.

$Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - The content of mitochondrial p62, OPTN, & VCP in spinal cord of SOD1‐G93A mice is decreased by Parkin knockoutImmunostaining of anterior horn of lumbar spinal cord for ChAT (in green) & p62 (in red) at 130 days. Scale bar, 10 μm.Quantification of the percentage of ChAT‐positive motor neurons (MN) containing p62‐positive inclusions at 130 days. PKO/G93A MN have less SOD1 aggregates than G93A, while Non Tg & PKO MN have none. Data were collected from n = 4 (two males & two females) mice for Non Tg & PKO & n = 6 (three males & three females) for G93A & PKO/G93A. Number of images recorded for each genotype: n = 9 for Non Tg & PKO, n = 16 for G93A, & n = 12 for PKO/G93A. Results are expressed as mean ± SEM; *P = 0.032, by unpaired one‐tailed Student's t‐test.Representative Western blots of OPTN, p62, & VCP in spinal cord mitochondrial fractions at 130 days. Protein levels are normalized by Complex V. The quantifications in panels (D–F) show that mitochondria of PKO/G93A mice have less mitophagy adaptor proteins than G93A relative to Complex V.Quantification of OPTN relative to Complex V at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); *P = 0.039 by paired Student's t‐test.Quantification of p62, relative to Complex V at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); **P = 0.0029 by paired Student's t‐test.VCP was quantified using Complex V as loading reference at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); *P = 0.044 by paired Student's t‐test.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30126943), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - G2019S LRRK2 hippocampal neurons do not have elevated alpha -synuclein pathology 14 days post-transduction. a Primary hippocampal neurons from NTG or G2019S pups were transduced with 2.5 μg/mL alpha -synuclein PFFs & allowed to age a further 14 days prior to sequential detergent fractionation. TX-100-insoluble alpha -synuclein & p62 are similar in both neuron types. b Quantification of soluble proteins shows ~ 25-fold elevation in the expression of LRRK2 & a commensurate ~ 50-fold elevation in pS395 LRRK2, indicative of the elevated LRRK2 kinase activity associated with the G2019S mutation. Soluble alpha -synuclein levels were equivalent between the cultures. c No significant differences were found between the genotypes in insoluble proteins by an unpaired t-test with Welch’s correction. (N = 3 biological replicates for each protein). Means + s.e.m.; **P < 0.01; *P < 0.05 by an unpaired t-test with Welch’s correction for unequal variances. All values are normalized to NTG neurons treated with alpha -synuclein PFFs & DMSO Image collected & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/s40478-018-0550-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - LRRK2 inhibition does not reduce insoluble alpha -synuclein in wildtype hippocampal neurons. a Primary hippocampal neurons from CD1 pups were treated with 30 nM LRRK2 inhibitors PF-475, PF-360 or DMSO as a vehicle control, then transduced with 2.5 μg/mL alpha -synuclein PFFs & allowed to age a further 14 days prior to sequential detergent fractionation. TX-100-insoluble alpha -synuclein & p62 are similar in both LRRK2 inhibitor treated & untreated neurons. b Quantification of soluble proteins show some reduction of LRRK2 protein levels PF-475 treatment & ~ 75% inhibition of LRRK2 activity (as assayed by pS935 LRRK2) by both PF-475 & PF-360. c Insoluble pS129 alpha -synuclein was slightly, but significantly elevated by PF-360 treatment, while alpha -synuclein & p62 were unchanged by one-way ANOVA. (N = 5 biological replicates for each protein). Means + s.e.m.; *P < 0.05, ****p < 0.0001 by one-way ANOVA with Dunnett’s multiple comparison test. All values are normalized first to GAPDH, as a loading control, then to DMSO-treated neurons Image collected & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/s40478-018-0550-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Colocalization of VANGL2 & p62/SQSTM1 in late endosomes.(a) Immunofluorescence staining of SKBR7 cells showed colocalization of endogenous VANGL2 (green) & p62/SQSTM1 (red) in discrete cytoplasmic puncta. Scale bar, 10 μm. The mean Pearson correlation for VANGL2 & p62/SQSTM1 is 0.62, calculated using the Image J software for ∼15 cells per field of view & from 10 images. (b) Partial colocalization of VANGL2 & p62/SQSTM1 in late endosomes of SKBR7 cells stained w/ the LAMP1 marker. Scale bar, 20 μm. (c) SKBR7 ells cultured in a nutrient-deprived medium & treated w/ 100 nM bafilomycin A1 (6 h) before fixation. Double labelling against VANGL2 (arrowheads) & p62/SQSTM1 (arrows), as described in Methods, showed accumulations of both markers in vesicular structures probably resembling endosomes/amphisomes. Scale bar, 200 nm. (d) IMCD3 cells treated w/ PBS or EGTA (5 mM) for 30 min. Immunofluorescence & confocal analysis performed using the indicated antibodies. Scale bar, 10 μm. Inserts show colocalized VANGL2 & p62/SQSTM1. (e) The VANGL2–p62/SQSTM1 complex recovered in confluent SKBR7 or IMCD3 cells w/ 2G4 mAb (IP VANGL2) but not a control antibody (IP IgG2a ctrl) as seen using western blot analysis w/ the indicated antibodies. The complex more abundant in cancer cells (SKBR7) than in polarized cells (IMCD3). Note that human (SKBR7 cells) & murine (IMCD3 cells) p62/SQSTM1 run at different molecular weights. (f) Lysates of confluent IMCD3 cells treated for 30 min w/ PBS or EGTA subjected to 2G4 mAb immunoprecipitation (IP VANGL2). Immunoprecipitated proteins probed by western blot analysis w/ the indicated antibodies. Increased amounts of VANGL2–p62/SQSTM1 complexes recovered after EGTA treatment. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms10318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Colocalization of VANGL2 & p62/SQSTM1 in late endosomes.(a) Immunofluorescence staining of SKBR7 cells showed colocalization of endogenous VANGL2 (green) & p62/SQSTM1 (red) in discrete cytoplasmic puncta. Scale bar, 10 μm. The mean Pearson correlation for VANGL2 & p62/SQSTM1 is 0.62, calculated using the Image J software for ∼15 cells per field of view & from 10 images. (b) Partial colocalization of VANGL2 & p62/SQSTM1 in late endosomes of SKBR7 cells stained w/ the LAMP1 marker. Scale bar, 20 μm. (c) SKBR7 ells cultured in a nutrient-deprived medium & treated w/ 100 nM bafilomycin A1 (6 h) before fixation. Double labelling against VANGL2 (arrowheads) & p62/SQSTM1 (arrows), as described in Methods, showed accumulations of both markers in vesicular structures probably resembling endosomes/amphisomes. Scale bar, 200 nm. (d) IMCD3 cells treated w/ PBS or EGTA (5 mM) for 30 min. Immunofluorescence & confocal analysis performed using the indicated antibodies. Scale bar, 10 μm. Inserts show colocalized VANGL2 & p62/SQSTM1. (e) The VANGL2–p62/SQSTM1 complex recovered in confluent SKBR7 or IMCD3 cells w/ 2G4 mAb (IP VANGL2) but not a control antibody (IP IgG2a ctrl) as seen using western blot analysis w/ the indicated antibodies. The complex more abundant in cancer cells (SKBR7) than in polarized cells (IMCD3). Note that human (SKBR7 cells) & murine (IMCD3 cells) p62/SQSTM1 run at different molecular weights. (f) Lysates of confluent IMCD3 cells treated for 30 min w/ PBS or EGTA subjected to 2G4 mAb immunoprecipitation (IP VANGL2). Immunoprecipitated proteins probed by western blot analysis w/ the indicated antibodies. Increased amounts of VANGL2–p62/SQSTM1 complexes recovered after EGTA treatment. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms10318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Parkin protein levels are unaffected in liver while a small decrease is measured in G93A cerebellumRepresentative Western blot for Parkin & p62 in cerebellum (Cb) homogenates at 130 days.Quantification indicates that Parkin is decreased in G93A mice relative to Non Tg at 130 days. Protein levels were normalized by beta ‐actin. Results are expressed as mean ± SEM & percent of Non Tg; n = 8 (four males & four females) mice per group. *P = 0.039 by paired Wilcoxon's test.Quantification of p62 in the homogenates indicates that p62 is increased in PKO & PKO/G93A mice, independent of SOD1‐G93A expression, at 130 days of age. beta ‐actin was used for normalization. Results are expressed as mean ± SEM & percent of Non Tg; n = 8 (four males & four females) mice per group. No statistically significant differences were found between Non Tg & G93A by paired Friedman's test with Dunn's correction (P = 0.99). ***P = 0.0001 by paired Student's t‐test (G93A vs. PKO/G93A).Representative Western blot of Parkin & p62 in liver homogenates at 130 days.Parkin protein levels were quantified at 130 days. beta ‐actin was used for normalization. Results are expressed as mean ± SEM & percent of Non Tg; n = 8 (four males & four females) mice per group. No statistically significant differences were found between Non Tg & G93A (P = 0.546 by paired Wilcoxon's test).Quantification of p62 protein levels in liver at 130 days. Results are expressed as mean ± SEM & percent of Non Tg; n = 8 (four males & four females) mice per group. No statistically significant differences were found between Non Tg & G93A (P = 0.546 by paired Wilcoxon's test). No statistically significant differences were found among the other groups.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30126943), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Parkin knockout mitigates the accumulation of mitophagy adaptors in G93A mitochondria at disease end stageA, BRepresentative Western blots of p62 (A), OPTN, & VCP (B) in spinal cord mitochondria from end‐stage mice.Cp62 quantification, using Complex V as normalizer, shows a strong accumulation of p62 in SOD1‐G93A mitochondria at end stage. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females) mice per group. No statistically significant differences were found between G93A & PKO/G93A (P = 0.078 by paired Wilcoxon's test); ***P = 0.0007 (Non Tg vs. G93A) & *P = 0.037 (PKO & PKO/G93A) both by paired Friedman's test with Dunn's correction.DQuantification of OPTN accumulation in end‐stage mitochondria, with Complex V as protein loading control. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females) mice per group. **P = 0.0078 (for G93A & PKO/G93A) by paired Wilcoxon's test; ***P = 0.0003 (Non Tg vs. G93A) by paired Friedman's test with Dunn's correction. No other statistically significant differences were found.EQuantification of VCP in mitochondria at disease end stage. Complex V was used as normalizer. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females) mice per group. *P = 0.039 (for G93A & PKO/G93A) by paired Wilcoxon's test; ***P = 0.0007 (Non Tg vs. G93A) & *P = 0.037 (PKO & PKO/G93A) both by paired Friedman's test with Dunn's correction.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30126943), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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AF835

Caspase-3 Antibody [Unconjugated] - Active

(7 Reviews)

(438 Publications)

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AF835

Caspase‑3 was detected in immersion fixed anti-FAS treated Jurkat human acute T cell leukemia cell line using 0.3 µg/mL Human/Mouse Active Caspase‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF835) for 3 hours at room temperature. Cells were stained (red) and counterstained (green). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Caspase-3 was detected in immersion fixed Jurkat human acute T cell leukemia cell line stimulated with staurosporin using Human/Mouse Active Caspase-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF835) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (yellow; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Caspase-3 was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using Rabbit Anti-Human/Mouse Active Caspase-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF835) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nucleus. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Representative sections of the medial side of right tibial epiphysis stained with/without caspase-3. Sections stained with caspase-3 antibody at four different time points (DH: A, C, E and G; BS2: B, D, F and H); a higher magnification (I); the growth plate (J) and fibrocartilage of anterior cruciate ligament (K) show caspase-3 positive cells (red cytoplasmic staining; arrow). A negative control section is shown in (L). Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/14/9/17729), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunohistochemical staining for caspase 3 on surgically transected tumours. A) Epo-treated tumour 24 hours after surgery. B) Placebo-treated tumour 24 hours after surgery. C) Placebo-treated tumour 48 hours after surgery Image collected and cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-14-648), licensed under a CC-BY license. Not internally tested by R&D Systems.

LPS challenge induces cell shedding from small intestinal villi. C57 BL/6 mice were administered either (a) PBS (control) or (b) LPS by IP injection and proximal small intestines removed after 1.5 h. Processed tissue was sectioned and stained by immunohistochemistry for CC3 (i.e. brown cells indicate shedding event), also highlighted by arrows. A representative picture for each group is shown (12 mice per group, two independent experiments). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28123052), licensed under a CC-BY license. Not internally tested by R&D Systems.

The cytoprotective effect of B. breve is MyD88 dependent. (a,b) IEC MyD88+/+ mice and (c,d) IEC MyD88−/− mice were gavaged with PBS (control) or B. breve UCC2003 and challenged with LPS. Paraffin-embedded intestinal sections were stained with anti-CC3 and quantified using the WinCrypts and Score programs. (e) Columns shown TLR2 expression via RT-PCR. Data are mean ± s.d., n = 12 (two independent experiments) analysed with Mann–Whitney U-test. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28123052), licensed under a CC-BY license. Not internally tested by R&D Systems.

Bifidobacterium breve UCC2003 protects against LPS-induced cell shedding. C57 BL/6 mice received three daily oral gavage doses of (a) PBS or (b) approximately 1 × 109B. breve UCC2003 followed by IP challenge with LPS 24 h later. Representative images are shown. Formalin-fixed, paraffin-embedded intestinal sections were sectioned and stained with anti-CC3 and (c) quantified using the WinCrypts and Score programs, 20 well-orientated hemi-villi were counted per mouse. Data are mean ± s.d., n = 12 (two independent experiments) analysed with a Mann–Whitney U-test. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28123052), licensed under a CC-BY license. Not internally tested by R&D Systems.

Genetic deletion of Bax prevents mutant LRRK2 induced neuronal death. Primary embryonic cortical neurons are derived from WT mice or mice lacking the pro-apoptotic protein Bax, and transiently transfected with GFP-tagged WT or mutant LRRK2 (R1441C, G2019S). (a) A representative image showing typical morphological features of apoptotic neurons expressing mutant LRRK2; that are positive for active caspase-3 (casp3) and display condensed and fragmented nuclei. (b) Quantification of apoptotic neuronal death (apoptotic nuclear profiles, “apo”; and active caspase-3 positive neurons, “casp3”) from cultures depicted in (a); **p < 0.01 in comparison to WT transfected neurons. (c) Primary neurons were prepared from WT mice or mice deficient in Bax, and treated with the topoisomerase I inhibitor camptothecin (“cam”, 5 μM, 18 hr), or doxorubicin (“doxo”, 10 ng/ml, 36 hr). Cells were fixed and processed for apoptotic nuclear counts. (d) Primary cortical neurons were prepared from WT mice or mice deficient in Bax, and transiently transfected with WT, R1441C, or G2019S LRRK2. As before, the cultures were fixed and the percentage of LRRK2-positive neurons that displayed apoptotic nuclear features was determined. The absence of Bax significantly protected neurons from death induced by mutant LRRK2. ***p < 0.001 compared to WT neurons expressing R1441C- or G2019S-LRRK2. Each data point represents the mean +/− SEM from 3–4 independent transfections from a representative culture. Cultures were repeated at least 3 times with similar differences. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29472595), licensed under a CC-BY license. Not internally tested by R&D Systems.

Hepatocyte ARNT deletion alters fibrotic and apoptotic gene expression in the TAA model of fibrosis.Results for FC are shown in black columns and LARNT in white columns. Results are expressed relative to FC level. (A) Messenger RNA expression of ARNT and collagen isotypes. (B) Expression of genes regulating fibrosis in FC and LARNT mice. (C) F4/80 and inflammatory cytokine expression in FC and LARNT mice. (D) Apoptotic gene expression in FC and LARNT mice. (E) Representative histology of FC liver stained with anti-Caspase 3 at 100X magnification. (F) Representative histology of LARNT liver stained with anti-Caspase 3 at 100X magnification. (G) Average Caspase 3 positive cell counts per field of view at 100 X magnification. Mean±SEM, * = p<0.05 ** = p<0.01, *** = p<0.001. N = 5-6/group. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0121650), licensed under a CC-BY license. Not internally tested by R&D Systems.

p140Cap expression is causative in limiting tumour growth in NeuT mice.(a) Expression cassette used for the generation of MMTV-p140Cap transgenic mice. The Myc epitope is inserted at the carboxyterminal region of the protein. (b) Extracts of tumours derived from NeuT mice (a,b) or p140-NeuT mice (c,d) were run on 6% SDS–PAGE and stained with antibodies to p140Cap, ERBB2 and tubulin for loading control. (c) Percentage of tumour free mice in NeuT (red line) and p140-NeuT (blue-dashed line) transgenic animals in both FVB (left) or BALB/c background (right). Twelve mice were analysed for each group. Fisher's exact test, Two sided, P=0,0022; P=0,0056. Error bar: s.e.m. (d) Total tumour burden in NeuT (red) and p140-NeuT (blue) mice in both the FVB (left) or BALB/c backgrounds (right) was measured. Ten mice were analysed in each group. Unpaired t test: (*P<0.05; ***P<0.001). Error bar: s.e.m. (e) Paraffin-embedded sections from three NeuT and three p140-NeuT tumours taken from mice at 33 weeks of age were analysed for hematoxylin–eosin (H&E) (a–d) and for immunohistochemistry with antibodies to NeuT (e–h), PCNA (i–l) and activated Caspase3 (m–p). Representative images are shown. Scale bar, 50 μm (first and third columns); 20μm (second and fourth columns). Histograms on the right show the percentage of PCNA+ (upper panel) and Activated Caspase3+ (lower panel) cells. Statistical significative differences were evaluated using unpaired t-tests (***P<0.001). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms14797), licensed under a CC-BY license. Not internally tested by R&D Systems.

Microglia-derived TNF-alpha alleviates aberrant neurogenesis.(a) qRT-PCR analyses of Tnf-alpha levels in the DG of WT and TLR9 KO mice at the indicated time points after seizure. Experimental controls were KA-untreated WT and TLR9 KO mice, respectively (n=3 animals). *P<0.05 by analysis of variance (ANOVA) with Tukey post-hoc tests. (b) Experimental scheme for assessing aNS/PC proliferation in microglia culture-derived CM pretreated with TNF-alpha -neutralizing antibody or IgG control. (c) Representative images of EdU (red), active caspase3 (green) and Hoechst (blue) staining in aNS/PCs cultured with the indicated CM from microglia (n=4 experiments). Scale bar, 50 μm. (d) Quantification of EdU+ or active caspase3+ cells in c (n=4 experiments). (e) Experimental timeline for assessing aNS/PC proliferation in thalidomide (Thal)-treated mice. (f) Representative images of BrdU+DCX+ newly generated immature neurons in the DG (n=4 animals). Scale bar, 50 μm. (g,h) Quantification of the number of BrdU+DCX+ cells in f, in the DG (g) and in the hilus (h) (n=4 animals). *P<0.05 and **P<0.01 by ANOVA with Tukey post-hoc tests. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25751136), licensed under a CC-BY license. Not internally tested by R&D Systems.

Double immunofluorescent staining of FSGS kidneys to Ki-67, alpha -tubulin and DAPI or double immunofluorescent staining to caspase-3 and DAPI. A. FSGS kidneys: cysts with simple squamous epithelium (CS) filled with colloid content, interstitium (i) and collecting tubules (ct) contain Ki-67-positive cells (arrows)(square). B. alpha -tubulin visualizes short cilia (arrows) in the cysts and extremely long cilia in collecting tubules (arrows)(square). C. Nuclear DAPI stain. D. Merging of A + B + C shows relationships between proliferating cells (green) and cilia (red). Immunostaining to Ki-67, alpha -tubulin and DAPI; scale bar 10 μm. E. Distribution of Ki-67 positive cells (%) in CNF and FSGS nephrotic syndrome. Cysts of proximal tubules with simple squamous epithelium (CS) or simple cuboidal epithelium (CC), proximal tubules with apparently normal simple cuboidal/columnar epithelium (PTNC). Data are shown as mean ± SD. Significant differences (Kruskal–Wallis) indicated by ***p < 0.0001. F. FSGS kidneys: caspase-3-positive apoptotic cells (arrows) in proximal tubules (PTNC) and interstitium (i). G. Nuclear DAPI stain. H. Merging of F + G shows green caspase-3- positive apoptotic nuclei and blue non-apoptotic nuclei. Immunostaining to caspase-3 and DAPI, scale bar 10 μm. Image collected and cropped by CiteAb from the following publication (//bmcnephrol.biomedcentral.com/articles/10.1186/1471-2369-15-3), licensed under a CC-BY license. Not internally tested by R&D Systems.

Epidermis-specific deletion of Rac1 increases UV-light-induced keratinocyte apoptosis in vivo. (a) H/E staining of UV-irradiated skin of Rac1 fl/fl and Rac1-EKO mice at 12 h after UV-irradiation. Black arrows indicate sunburn cells. (b) Graph shows the percentage number of sunburn cells at 12 h with (red bars) or without (blue bars) UV-irradiation in Rac1 fl/fl (n=4) and Rac1-EKO (n=5) mice. The percentage of sunburn cells within the epidermis after UV-irradiation in Rac1 fl/fl mice was 3.4%, whereas in Rac1-EKO mice it was 8.6% of total epidermal keratinocytes. Non-irradiated samples showed <1% sunburn cells in both the genotypes. (c) Immunostainings against cleaved caspase-3 (green) of UV-irradiated skin of Rac1 fl/fl and Rac1-EKO mice at 12 h after UV-irradiation. Nuclei are stained in blue. Scale bar=100 μm. (d) Western blot analysis of cleaved caspase-3 from epidermal lysates of untreated (no UV) and UV-light treated (UV) Rac1 fl/fl and Rac1-EKO mice. Non-irradiated controls showed no bands for cleaved capsase-3, whereas samples of irradiated epidermis showed cleaved caspase-3-specific bands at ~25 kDa and 23 kDa. Numbers on the left denote molecular weights in kDa. (e) Graph shows densitometry analysis of western blot in (d). Error bars show S.D. Asterisks show P-value<0.001 Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28277539), licensed under a CC-BY license. Not internally tested by R&D Systems.

DNA damage has an important role in increased UV-light-induced keratinocyte apoptosis in Rac1-EKO epidermis in vivo. (a and c) H/E stainings and immunostainings against cleaved caspase-3 (green) of UV-irradiated skin of CPDPL/Rac1-EKO mice kept in the dark and under the photoreactivation lamp. Nuclei are stained in blue. Dashed line in c demarcates border between epidermis and dermis. Scale bar=100 μm. (e) Western blot analysis of cleaved caspase-3 from epidermal lysates of UV-irradiated CPDPL/Rac1-EKO mice kept in the dark and under the photoreactivation lamp. GAPDH was used as a loading control. Numbers on the left denote molecular weight in kDa. (b,d, and f) Graphs show quantification of sunburn cells (n=5, 4) (b), cleaved caspase-3 positive cells (n=9, 8) (d) and densitometric analysis of cleaved caspase-3 western blots (n=4, 6) (f) from UV-irradiated CPDPL/Rac1-EKO mice kept in the dark (black bars) and under the photoreactivation (PR) lamp (gray bars). Error bars show S.D. Asterisks show P-value. **<0.01, ***<0.001 Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28277539), licensed under a CC-BY license. Not internally tested by R&D Systems.

Smad1 deletion reduces oxidative stress and apoptosis following stroke.(A) Fluorescent images and quantification of in vivo HEt assay reflecting ROS levels on the ipsilateral hemisphere at 7 days post-stroke. The extent of oxidized nucleotides was measured by IHC for 8-oxo-dG levels at 24 h post-stroke. (B) IHC and quantification of activated Caspase 3 (aCasp3) and p53 on ipsilateral hemisphere at 7 days post-stroke. n = 3 for HEt, 8-oxo-dG, aCasp3, n = 4 for p53, unpaired Student’s t-test, *p<0.05; ***p<0.001. Scale, 50 μm. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0136967), licensed under a CC-BY license. Not internally tested by R&D Systems.

Effects of IGF-1 or/and PDGF-bb on IL-1 beta -induced NF-kappa B-dependent pro-inflammatory, pro-apoptotic and matrix degrading gene products in chondrocytes.To determine whether IGF-1 or/and PDGF-bbexert effects on IL-1 beta -induced NF-kappa B-dependent expression of pro-inflammatory, pro-apoptotic and matrix degrading gene products, primary chondrocytes were either stimulated with 10 ng/ml IL-1 beta , 10 ng/ml PDGF-bb, 10 ng/ml IGF-1 or combination of both growth factors (5 ng/ml each) or pre-stimulated for 12 h with 10 ng/ml PDGF-bb, 10 ng/ml IGF-1 or combination of both growth factors (5 ng/ml each) followed by 10 ng/ml IL-1 beta for 24. Equal amounts of total proteins were separated by SDS-PAGE and analyzed by immunoblotting using antibodies raised against COX-2, MMP-9 and MMP-13 and active caspase-3. Stimulation with IL-1 beta resulted in production of COX-2, MMP-9, MMP-13 and caspase-3 cleavage. Pre-treatment with a combination of both IGF-1 or/and PDGF-bb downregulated COX-2, MMP-9, MMP-13 and cleaved caspase-3. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0028663), licensed under a CC-BY license. Not internally tested by R&D Systems.

Allelic loss of Cdh11 led to faster growing tumors due to decreased cell death. (E) Cadherin-11 was knocked down using stealth siRNA in a cadherin-11 expressing cell line derived from TAg-RB tumors, T +539. Knockdown of Cdh11 clearly decreased caspase-3 expression compared to control. Image collected & cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/20421947), licensed under a CC-BY license. Not internally tested by R&D Systems.

Curcumin inhibits IL-1 beta activity, enabling co-culture induced chondrogenesis in MSCs. A: Fourteen days high-density culture. Untreated MSC cultures became apoptotic (a). In primary chondrocyte cultures (b), co-cultures (c), co-cultures treated with curcumin (e) or co-cultures pre-stimulated four hours with curcumin (f), prominent chondrogenesis was observed. Stimulation of the co-culture with IL-1 beta alone resulted in degeneration of the cell culture (d). In contrast, a four-hour pre-stimulation of the co-culture with curcumin followed by IL-1 beta incubation (g) or a four-hour pre-stimulation of the co-culture with curcumin followed by IL-1 beta and curcumin incubation (h) inhibited the adverse effects of IL-1 beta on the chondrogenic potential of the co-culture and prominent chondrogenesis was observed. Magnification, 6,000×; bar, 1 μm; C, chondrocytes, F, fibroblast-like cells; M, ECM. B-D: Immunoblots of whole cell lysates were probed with antibodies against CSPGs, collagen type II, beta 1-integrin, Shc, activated-ERK1/2, Sox-9, activated-caspase-3 and COX-2. In co-cultures pre-stimulated for four hours with curcumin followed either by incubation with IL-1 beta alone or incubation with IL-1 beta and curcumin, prominent production of chondrogenic matrix and adhesion molecules (B), activation of the chondrogenic signalling pathway (C) and down-regulation of apoptotic and inflammatory markers (D) was observed. Each experiment was performed in triplicate. Expression of the housekeeping gene beta -actin was not affected. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/20594343), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

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,

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Rabbit Polyclonal

Host: Rabbit Polyclonal

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Carrier Free	AF835-SP

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KGE004B

Prostaglandin E2 Parameter Assay Kit

(444 Publications)

1 Kit

KGE004B

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All-Multi

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Biotin	SKGE004B
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NB400-105

ABCA1 Antibody - BSA Free

(7 Reviews)

(419 Publications)

0.1 ml

NB400-105

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Reactivity:

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

Species Glossary

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

Applications:

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

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

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-105AF350
Alexa Fluor 405	NB400-105AF405
Alexa Fluor 488	NB400-105AF488
Alexa Fluor 532	NB400-105AF532
Alexa Fluor 594	NB400-105AF594
Alexa Fluor 647	NB400-105AF647
Alexa Fluor 700	NB400-105AF700
Alexa Fluor 750	NB400-105AF750
Alexa Fluor Plus 405	NB400-105AFP405
Alexa Fluor Plus 488	NB400-105AFP488
Alexa Fluor Plus 555	NB400-105AFP555
Alexa Fluor Plus 594	NB400-105AFP594
Alexa Fluor Plus 647	NB400-105AFP647
Alexa Fluor Plus 680	NB400-105AFP680
Allophycocyanin	NB400-105APC
Allophycocyanin/Cy7	NB400-105APCCY7
Biotin	NB400-105B
CoraFluor 1	NB400-105CL1
DyLight 350	NB400-105UV
DyLight 405	NB400-105V
DyLight 488	NB400-105G
DyLight 550	NB400-105R
DyLight 594	NB400-105DL594
DyLight 650	NB400-105C
DyLight 680	NB400-105FR
DyLight 755	NB400-105IR
FITC	NB400-105F
HRP	NB400-105H
Janelia Fluor 525	NB400-105JF525
Janelia Fluor 549	NB400-105JF549
Janelia Fluor 646	NB400-105JF646
Janelia Fluor 669	NB400-105JF669
PE	NB400-105PE
PE/Atto594	NB400-105PEATT594
PE/Cy5.5	NB400-105PECY55
PE/Cy7	NB400-105PECY7
PerCP	NB400-105PCP
mFluor Violet 450 SE	NB400-105MFV450
mFluor Violet 500 SE	NB400-105MFV500
mFluor Violet 610 SE	NB400-105MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-105SS

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MAB002

Mouse IgG1 Isotype Control (11711) [Unconjugated]

(1 Review)

(431 Publications)

500 μg

MAB002

Human peripheral blood mononuclear cells (PBMCs) werestained with Mouse Anti-Human LILRA6/CD85b Monoclonal Antibody (Catalog #MAB8656,filled histogram) or Mouse IgG1 Isotype Control Antibody (Catalog # MAB002, openhistogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG SecondaryAntibody (Catalog # F0101B).

EC and PC ICAM-1 expression and effects of CD18 blocking on neutrophil adhesion.ICAM-1 expression on (A) EC or (D) PC monolayers before and after 4 hr IL-1 beta activation was assessed by flow cytometry (IgG labeled control white). ICAM-1 expression on (B,C) EC and (E,F) PC following 4 hr IL-1 beta activation was imaged using confocal microscopy. Scale bars are 20 µm. (G) Neutrophil adhesion inhibition to IL-1 beta activated EC or PC monolayers. Freshly isolated neutrophils were pre-incubated with anti-Mac-1 or anti-LFA-1 antibodies and seeded onto EC or PC monolayers (activated for 4 with IL-1 beta ) in Sykes-Moore chambers and allowed to adhere prior to counting. Bars represent average neutrophil adhesion ± SEM. *P<0.05, when compared to the no block control. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0060025), licensed under a CC-BY license. Not internally tested by R&D Systems.

miR-720 maintains the migratory and invasive properties of MDA-MB-231 cells. MDA-MB-231 cells were transfected with 50 nM anti-miR-720 or anti-miR-negative control (Ctrl) for 48 h. Either RNA was isolated (a) or transfected cells subjected to functional assays (b–e). a miR-720 levels. RNA was subjected to RT-qPCR for miR-720. Control condition (Ctrl) was set to 1.0 (mean ± SD from three independent experiments). b Migration assay. Transfected cells were subjected to a migration assay in a Boyden chamber over a 24-h period. The cells that migrated through the chamber were quantified using crystal violet staining. Ctrl was set to 100 % (mean ± SD from three independent experiments). c Matrigel assay. Transfected cells were grown in Matrigel for 11–15 days and then photographed using a Nikon eclipse TS100 microscope at 10× magnification. Scale bar = 100 μm. Representative images of two independent experiments with similar results are shown (left panels). The number of branched colonies were counted. The count in the control sample was set to 1.0 (right panel). d Transendothelial migration assay. Transfected MDA-MB-231 cells were plated in a Boyden chamber that had been coated with a confluent monolayer of HUVEC cells, and the cancer cells that invaded through the HUVEC layer over a 24-h period were quantified using crystal violet staining. Ctrl value for transendothelial (transendo.) migration was set to 100 % (mean ± SD from three independent experiments) and relative (Rel.) migration values given. e ATP assay. Transfected cells were grown for 24 h and ATP levels were measured using an ATPlite 1Step assay. Ctrl was set to 100 % (mean ± SD from three independent experiments). *P < 0.05, ***P < 0.001, Student’s t test. Not sig. not significant Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27039296), licensed under a CC-BY license. Not internally tested by R&D Systems.

Monoclonal ASM antibody (23A12C3) inhibits pathogenic Th17 cell differentiation and protects neuropathological features in APP/PS1 mice. e Representative flow cytometry plot and graph displaying the calculated percentage and mean fluorescent intensity of Th17 cells (CD4+ IL17A+) differentiated from CD4+ T cell in each group (Control, n = 5; Smpd1ox/ox serum/con IgG ab, n = 10; Smpd1ox/ox serum/23A12C3, n = 8). f Western blotting for p-Stat3, p-JNK, p-AKT, p-mTOR in Th17 cells differentiated from CD4+ T cell in each group (n = 4/group). g ASM activity in plasma (n = 5/group). h Percentage of Th17 cells in the spleen, blood, and brain by flow cytometry (n = 5/group). i Quantification of microglia (Iba-1) and astrocyte (GFAP) (n = 5/group). j mRNA levels of inflammatory markers in the cortex and hippocampus (n = 4/group). k Representative immunofluorescence images and quantification of ThioS (A beta plaques) in cortex and hippocampus. Scale bars, 50 μm. (n = 5/group). l Quantification of 6E10 (n = 5/group). m The results of Morris water maze test (n = 10/group). n, o At probe trial day 11, time spent in target platform (n) and the number of times each animal entered the small target zone (o) were analyzed (n = 10/group). p The results of fear-conditioning test (n = 10/group). d–j, m–p One-way analysis of variance, Tukey’s post hoc test. k, l, n Two-tailed student’s t test. All error bars indicate s.e.m. All in vivo data analysis was done at 9-mo-old mice. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/36959217), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

Applications:

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 405	IC002V
Alexa Fluor 488	IC002G
Alexa Fluor 594	IC002T
Alexa Fluor 647	IC002R
Alexa Fluor 700	IC002N
Alexa Fluor 750	IC002S
Allophycocyanin	IC002A
Biotin	IC002B
Fluorescein	IC002F
PerCP (Peridinin-chlorophyll Protein Complex)	IC002C
Phycoerythrin	IC002P

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	IC002A
Carrier Free	IC002B
Carrier Free	IC002C
Carrier Free	IC002F
Carrier Free	IC002G
Carrier Free	IC002N
Carrier Free	IC002P
Carrier Free	IC002S
Carrier Free	IC002T
Carrier Free	IC002R
Carrier Free	IC002V

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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

Inhibitor Family

Inhibitor Target

Isotype

Kit Type

Lysate Life Stage

Lysate Protein State

Lysate Subcellular Fraction

Lysate Tissue

Lysate Type

Preservative

Protein Or Peptide

Lysate Tissue Condition

Research Areas

Has Sample Size

Promotional Item

Species Glossary