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NB100-2323

IRE1 alpha [p Ser724] Antibody - BSA Free

(9 Reviews)

(335 Publications)

0.1 ml

NB100-2323

P-Ire1a activation correlates with WFS1 cell expression. Cryosections of retina from 12 month old Wfs1+/+ and Wfs1-/- mouse were immunostained with anti-P-Ire1a antibody (red). DAPI was used for staining of cell nuclei (blue). RPE, retinal pigment epithelium; ONL, outer nuclear layer; INL, inner nuclear layer, RGC, retinal ganglion cells. Scale bars=50 um. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0097222) licensed under a CC-BY license.

Hepatic ER stress markers with cachexia progression. ER stress markers Bip, IRE1a, ATF6p50 and CHOP were examined in the liver of non, pre and severely cachectic mice. (n = 6-8 per group, p < 0.05) Dotted line indicates levels of Non-cachectic mice. Non = Non-Cachectic Apc Min/+ Sev = severely cachectic Apc Min/+; * denotes significantly different from Non-cachectic Apc Min/+ Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0119888) licensed under a CC-BY license.

IRF-1 plays a central role in ER stress-mediated modulation of VCAM-1 expression by TGRL. HAEC were conditioned for 4 hr with TNFa (0.3 ng/ml) E: CHOP knockdown decreased TNFa-induced VCAM-1 expression. n=4. **P<0.01 vs. siCtrl+TNFa. Shown are representative blots from 3 independent experiments with similar results. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0078322) licensed under a CC-BY license.

Analysis using HRP conjugate of NB100-2323. Detection of phosphorylated IRE-1 alpha using NB100-2323. Lane 1: COS-7 untransfected Lane 2: COS-7 expressing wild-type IRE1 alpha Lane 3: COS-7 expressing kinase-dead IRE1 alpha. Theoretical molecular weight: 110 kDa.

<b>Biological Strategies Validation. </b>Analysis of anti-IRE1 alpha (pSer724) using Lot AN and AO of NB100-2323. HeLa cells were treated (+) or untreated (-) with 10 mM DTT for 60 min to activate the UPR. Total protein was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% BSA in TBST. The membrane was probed with 2.0 ug/ml antibody in 5% BSA, and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Theoretical molecular weight: 110 kDa.

Regulation of the unfolded protein response by KLF15. Western analysis of UPR activity in WT versus KLF15-/- primary hepatocytes. Hepatocytes were isolated from standard chow-fed 4-month-old male WT and KLF15 -/- mice. Two individual experiments were performed in triplicate; each lane indicates a technical replicate. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0077851) licensed under a CC-BY license.

HeLa cell lysate, 20 ug. Antibody at 1:1000, 1% skim milk in TBST, overnight incubation at 4C. WB image submitted by a verified customer review.

<b>Biological Strategies Validation. </b>Detection in Min6 cells which were treated with different concentrations of glucose for 3 hours prior to lysates preparation.

IRE1 (pS724) was detected in immersion fixed paraffin-embedded sections of human spleen using Rabbit Anti-Human IRE1 (pS724) polyclonal Antibody (Catalog # NB100-2323) at 1:300 for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the perinuclear cytoplasm in splenocytes.

Hepatic ER Stress.Representative hepatic western blots of BiP, phosphorylated (Ser51), total EIF2 alpha and phosphorylated:total EIF2 alpha , phosphorylated (Ser724), total IRE1 alpha and phosphorylated:total IRE1 alpha , and CHOP (n = 9). Diets not sharing a common letter differ significantly from one another (P≤.05).

Morphine suppressed 6-OHDA-induced ER stress through activation of UPR. (A,B) Morphine induced UPR in SH-SY5Y cells. Protein levels of GRP78, p-IRE1 alpha , IRE1 alpha , p-PERK, PERK, ATF6, CHOP and Tubulin in SH-SY5Y cells were analyzed (A) and quantified (B) by western blot. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 vs. control.

Effect of cancer on ER stress markers.Bip1, IRE-1, ATF-6 p50 and CHOP expression in the liver of non—cachectic ApcMin/+ mice (N = 6 per group), compared to healthy C57BL/6 mice. Dotted line on the western blot indicates two different sections of the same gel. Values are expressed as Mean ± SE. * denotes significantly different from the healthy C57BL/6 mice as analyzed by a pre—planned t—test. p < 0.05.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - ABT-263 induces ER stress & activation of the unfolded protein response.a Western blot of ER stress molecules in uveal melanoma cells treated with ABT-263 5 μM for 24 h & 48 h. HSP90 is used as a loading control. Representative western blots are shown. b XBP1 splicing in uveal melanoma cells exposed to ABT-263 5 μM for 15 h. *P-value < 0.05, **P-value < 0.01. c Kinetic analysis of ER stress proteins in Mel270 cells treated with ABT-263 5 μM in the absence or presence of qVD-OPh 20 μM. HSP90 is used as a loading control. Representative western blots are shown. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32337074), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Effect of cancer on ER stress markers.Bip1, IRE-1, ATF-6 p50 & CHOP expression in the liver of non—cachectic ApcMin/+ mice (N = 6 per group), compared to healthy C57BL/6 mice. Dotted line on the western blot indicates two different sections of the same gel. Values are expressed as Mean ± SE. * denotes significantly different from the healthy C57BL/6 mice as analyzed by a pre—planned t—test. p < 0.05. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0119888), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - ER stress is induced by acidosis activated OGR1-mediated signalling. Caco-2 cells subjected to different pH medium, following 4–6 h incubation in pH 7.6 serum free medium.(B) After 24 h pH shift, total protein was isolated & WB was performed. The results representative of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31996710), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Expression of ER stress markers in mouse & human skeletal muscle(a) TA muscle lysates from high fat fed mice were used to determine the expression of ER stress markers by Western blot analysis (n=6). (b-g) Western blot analysis was done to measure proteins involved in ER stress in vastus lateralis muscle from lean, obese, or diabetic subjects (n=6-13). Data are the means ± S.E.M. * indicates p<0.05 & ** indicates p<0.01 vs. chow or lean controls, using Student’s t-test (a) & One way ANOVA. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms2851), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - The inositol-requiring enzyme 1 alpha (IRE1 alpha )/ X-box-binding-protein 1 spliced isoform (XBP1s) arm of unfolded protein response (UPR) is induced by angiotensin II in VSMCs. (A–C) The rat aortic VSMCs infected with adenovirus encoding GRP78 or control GFP (100 moi) for 48 h were stimulated with 100 nM AngII (AII) for 1–6 h & immunoblotting was performed as indicated. (A) Representative blots from 4 independent experiments. (B) Signal intensity was used to calculate the expression ratio of XBP1s to GAPDH. (C) Signal intensity was used to calculate the IRE1 alpha Ser724 phosphorylation ratio to the total IRE1 alpha . The bars in the graphs show the mean ± SD from 4 independent experiments. * indicates p < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32679678), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Morphine suppressed 6-OHDA-induced ER stress through activation of UPR. (A,B) Morphine induced UPR in SH-SY5Y cells. Protein levels of GRP78, p-IRE1 alpha , IRE1 alpha , p-PERK, PERK, ATF6, CHOP & Tubulin in SH-SY5Y cells were analyzed (A) & quantified (B) by western blot. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 vs. control. (C,D) Protein levels of GRP78, CHOP & Actin in SH-SY5Y cells were analyzed (C) & quantified (D) by western blot in the indicated groups. ∗P < 0.05, ∗∗P < 0.01 vs. control & #P < 0.05, ##P < 0.01 vs. 6-OHDA-treated group. All data are expressed as mean ± SEM. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fnmol.2018.00120/full), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Hepatic ER Stress.Representative hepatic western blots of BiP, phosphorylated (Ser51), total EIF2 alpha & phosphorylated:total EIF2 alpha , phosphorylated (Ser724), total IRE1 alpha & phosphorylated:total IRE1 alpha , & CHOP (n = 9). Diets not sharing a common letter differ significantly from one another (P≤.05). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0167979), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Effect of Insig-1 overexpression on nSREBP-1c & proteins related to ER stress pathway in INS-1 cells. INS-1 & INS-1-Insig-1 cells were stimulated with 11.2 mM & 25.0 mM glucose for 0, 24 & 72 h. A) nSREBP-1c & p-IRE1 alpha pathway of ER stress related proteins were detected by Western blot. GAPDH was used as control for protein loading. Equal amount of proteins (50 μg) were loaded. B) Relative protein expression compared to GAPDH, data are the means ± SE of three independent experiments; *, P < 0.05, **, P < 0.01 VS INS-1 cells at the same time point. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21843373), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - cCK-18 is formed following activation of the UPR. A. Immunoblots for phospho-IRE1 (p-IRE1), total IRE1 (IRE1), & cleaved cytokeratin 18 (cCK-18) showing time-dependent increase of cCK-18 in A549 cells exposed to 0.5 μM of thapsigargin (TG, +) for various lengths of time. Immunoblots showing an increase in total & phospho-IRE1 confirm thapsigargin activates the UPR in A549 cells compared to vehicle controls (−). B. Immunoblots for cCK-18 showing time-dependent appearance of cCK-18 in annexin V– or V+ A549 cells exposed to 0.5 μM of thapsigargin for various lengths of time. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23167970), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Protein expression of downstream markers of ER stress in islets from lean, obese & high-fat-diet-fed fZDF rats. Protein lysates from islets isolated from three age-matched fZDF rats (obese), HF-fZDF (HFD) rats & their heterozygous lean littermates (lean) were separated by SDS-PAGE & analysed by western blotting. As controls, lysates from the islets isolated from Wistar rats or MIN6 cells treated with 1 μM thapsigargin for 2 h or sorbitol for 1 h were run alongside. Proteins were detected using antisera against (a) Calreticulin, HYOU1, GRP78, p-IRE1 alpha , p-eIF2 alpha & CHOP & as protein loading control rpS6 & (b) p-JNK & total JNK1/2 as loading control. Western blots were quantified using Image-J software. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/nutd201335), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - CETP inhibition reduces endoplasmic reticulum stress. HAECs transfected with CETP or control scrambled siRNA. (A) Western blot analysis of PERK (n = 3, * p < 0.01) & qPCR analysis of ATF6 mRNA. (B) Phosphorylated & total IRE (n = 3, ns). (C) Representative microscopic images of HAECs stained with GADD153, n = 5, & (D) Grp78/BIP, & DAPI. Scale bar, 10 μm. Mean ± SEM, n = 5, * p < 0.05. HAECs, human aortic endothelial cells. CETP, cholesteryl ester transfer protein. NS, non-silenced. siCETP, silenced CETP. PERK, protein kinase RNA-like ER kinase. ATF6, Activating transcription factor 6. IRE, inhibitor resistant esterase. BIP, binding immunoglobulin protein. GADD153, growth arrest- & DNA damage-inducible gene 153. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33430172), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Activation of UPR signaling pathways after UIRI. a & b The expression of p-PERK & t-PERK in kidneys of UIRI mice were evaluated with western blot analysis & quantified. c UIRI induces the activation of ATF6 as demonstrated by enhanced expression of cleaved ATF6 (cATF6). GAPDH was used as an internal control. d Fold-change expression of cATF6 in kidneys of UIRI mouse as compared with that of sham group mouse. e & f The expression of p-IRE1 & t-IRE1 in mice kidneys were evaluated with western blotting & quantified. Data are expressed as means ± SEM, n = 3 ~ 6 in each group. *P < 0.05, **P < 0.01, & ***P < 0.001, as compared with sham group Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35765067), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Apoptosis mediated by Arl6ip5 interference is regulated by Chop. (a) Immunoblotting was used to detect the protein level for p-IRE1 alpha , IRE1, Chop, Arl6ip5 & beta -actin in the UAMS-32 cell with NC-siRNA or Arl6ip5-siRNA treatment for 72 h. Representative results were shown from three independent experiments. (b–e) UAMS-32 cells first received NC-siRNA or Chop-siRNA for 24 h, then the medium was changed & further received NC-siRNA or Arl6ip5-siRNA for 72 h. Q-PCR was used to analyze the mRNA levels of Chop (b), GADD34 (c) & Trib3 (d). The apoptotic cells were stained with Annexin-V & the proportion were calculated (e). n=4. In all panels, error bars represent the mean±S.E.M. **P<0.01, *P<0.05 based on ANOVA Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014427), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Downregulation of gamma -taxilin & alpha NAC, & ER stress responses in mouse brain slice cultured under hypoxic conditions. (a) Immunohistochemistry shows downregulation of gamma -taxilin & alpha NAC expression in brain slices cultured for 48 h under hypoxic conditions. Scale bar, 20 μm. (b) Western blot shows downregulation of gamma -taxilin & alpha NAC, phosphorylation of PERK & IRE1 alpha & induction of CHOP in mouse brain slices cultured for 48 h under normoxic (−) or hypoxic conditions (+) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25880086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Differential ER stress response pathways initiated by gamma -taxilin or alpha NAC depletion. Proteins involved in UPR sensor (PERK, IRE1 alpha ), eIF2 alpha , JNK, Bcl-2, & Bcl-2-related protein (Puma, Bim, Bak, & Bax) signaling pathways were analyzed by Western blotting of HeLa S3 cells that were depleted of gamma -taxilin or alpha NAC by RNA intereference for 72 & 96 h. Upper panels, taxilin & NAC proteins; middle panels, UPR proteins; lower panels; apoptotic proteins; & bottom panel, beta -actin Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25880086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - The UPR is activated in type II AECs isolated from IPF lungs. Phosphorylated IRE-1 alpha (panel A) & spliced XBP-1 (panel B) were present in increased amounts in AECs from 3 different IPF lungs, but not in the 3 control subjects (* P < 0.05, **P = 0.0495 by Wilcoxon rank sum test). # indicates amplicon of unspliced XBP1 digested with Pst1. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23167970), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - ER stress is induced by OGR1 via IRE1 alpha /JNK signalling. (A) Caco-2 cells were subjected to different pH medium, with or without an OGR1 inhibitor (10 µM), following 4–6 h in pH 7.6 serum free medium. After 24 h pH shift, total protein was isolated & Western blot performed. Results are representative of two independent experiments. (B) Caco-2 cells were subjected to different pH medium After 24 h pH shift, total protein was isolated & Western blot performed. Results are representative of two independent experiments. (C) Caco-2 cells were subjected to different pH medium with or without a JNK inhibitor (10 µM) following 4–6 h in pH 7.6 serum free medium. After 24 h pH shift, total protein was isolated & Western blot performed. Results are representative of two independent experiments. (D) Caco-2 cells were starved & subjected to an acidic pH with or without a JNK inhibitor as described in (C). After 24 h pH shift, total RNA was isolated & mRNA expression was investigated by qPCR. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post-test. Data are presented as means ± SE of three independent experiments (***p < 0.001). (E) Caco-2 cells were starved & subjected to different pH medium following 4–6 h in pH 7.6 serum free medium. After 24 h pH shift, total protein was isolated & co-IP using IRE1 alpha antibody & JNK antibody was performed, followed by immunoblotting. Results are representative of two independent experiments. pH conditions: High pH 7.5–7.8; Normal pH 7.2–7.4; Low pH 6.6–6.8. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31996710), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Downregulation of gamma -taxilin & alpha NAC & ER stress responses in hypoxic cells. (a) Phase-contrast micrographs of SK-N-SH cells cultures in hypoxic conditions for 0–48 h. Floating cells were evident by 37 h after the beginning of hypoxic treatment. (b) FACS (fluorescence-activated cell sorting) analysis of apoptosis in hypoxic SK-N-SH cells. Bars in the FACS profiles (left panel) indicate the fraction locations of annexin-positive cells. Bar graph indicates the relative numbers of annexin-positive cells (right panel). Bar graph data are shown as means±S.D. (n=3). *Significantly different from controls (0 h) (P<0.001, Tukey–Kramer test). (c) DAPI (4',6-diamidino-2-phenylindole) staining shows fragmented nuclei of apoptotic SK-N-SH cells after hypoxic treatment. Scale bar, 10 μm. (d) Downregulation of gamma -taxilin & alpha NAC & induction of ER stress responses in hypoxic SK-N-SH cells. Western blot analysis was performed 0 to 48 h after hypoxic treatment of cells. Upper panels, taxilin & NAC proteins; middle panels, UPR proteins; lower panels, apoptotic proteins; & bottom panels, cleaved caspase-9 & beta -actin proteins. (e) MG-132 treatment restore the gamma -taxilin & alpha NAC protein level in hypoxic (16 h) HeLa S3 cells. MG-132 (−), DMSO; MG-132 (+), 20 μM. (f) Caspase inhibition does not affect the hypoxia-induced downregulation of gamma -taxilin & alpha NAC in HeLa S3 cells. (g) Colocalization of gamma -taxilin & alpha NAC in normoxic (0 h) & hypoxic (8 h) HeLa S3 cells. Note that the colocalization fraction increases with the appearance of apoptotic phenotypes. Scale bars, 10 μm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25880086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - gamma -Taxilin ablation causes ER stress responses & apoptosis in normoxic cells. (a) Knockdown of gamma -taxilin expression by small interfering RNA (siRNA) in HeLa S3 cells. Two different siRNA constructs are equally effective. HeLa S3 cells were treated with solvent alone (Mock), control siRNA (Co), or gamma -taxilin siRNA ( gamma -tax-1 or -2) for 96 h. (b) Induction of apoptosis by gamma -taxilin ablation in HeLa S3 cells. gamma -Taxilin ablation causes apoptosis in HeLa S3 cells, but Mock or Co treatment did not. Bar graph shows fractions of annexin-positive cells (means±S.D., n=3). *Significantly different from Mock- or Co treatment (P <0.001, Tukey–Kramer test). (c) Confocal microscopy demonstrates coincidence of gamma -taxilin depletion & apoptotic nuclei in HeLa S3 cells treated with gamma -taxilin siRNA. Scale bar, 10 μm. (d) gamma -Taxilin depletion triggers ER stress responses in HeLa S3 cells. Upper panels, taxilin & NAC proteins; middle panels, UPR proteins; & lower panel, beta -actin. (e) gamma -Taxilin ablation induces accumulation of ubiquitinated proteins in HeLa S3 cells. Cell lysates were analyzed on 7.5% SDS-PAGE, followed by immunoblotting with antibodies specific for anti-mono- & polyubiquitinated conjugates (upper panel) & beta -actin (lower panel) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25880086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Localization of the WT & mutant Wrn proteins in different subcellular fractions.(A) Example of western blots showing the presence of the Wrn WT protein in the nuclear fraction & the presence of the Wrn delta hel mutant protein mainly in the cytosolic fraction. Fractionations were performed on WT, Wrn delta hel/ delta hel, & Wrn-/- MEFs. Topoisomerase I & beta -tubulin are nuclear & cytoplasmic markers, respectively. As expected the molecular weight of the Wrn delta hel mutant protein was lower than the Wrn WT protein. No band was detected in Wrn-/- MEFs fractions with the anti-Wrn polyclonal antibody. (B) Example of western blots showing only the presence of the Wrn delta hel mutant protein in total ER & the peroxisomal fractions. Fractionations were performed on WT, Wrn delta hel/ delta hel, & Wrn-/- liver tissues. Catalase & calreticulin are used as peroxisomal & ER specific markers. This experiment was repeated three times. (C) Protein levels of GRP78, phosphorylated IRE alpha , & beta -actin (top panels) & of total IRE alpha , & beta -actin (bottom panels) in the spleen of three animals of each genotype. (D) Ratio of GRP78 signal over beta -actin signal from western blots. (E) Ratio of IRE alpha over beta -actin signal from western blots. (F) Ratio of phosphorylated IRE alpha signal over beta -actin signal from western blots. Experiments were performed in triplicates. Bars in all histograms represent SEM. Tukey post ANOVA test P-values are shown (*P < 0.05 & **P < 0.01). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26447695), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - IER3IP1 suppression decreases the UPR activation(A) mRNA levels of IRE1 alpha , Perk & Atf6 in IER3IP1 KD cells. 4 days after infection with IER3IP1 shRNA lentivirus, mRNA levels of IRE1 alpha , Perk & Atf6 in IER3IP1 KD cells were determined by qRT-PCR. *P<0.05, **P<0.01 compared to control group. Values are mean ± SEM. (B) Effect of IER3IP1 suppression on the IRE1 alpha arm of the UPR. 4 days after IER3IP1 KD in MIN6 cells, phosphorylation of IRE1 alpha , Bip, sXBP-1 & uXBP-were determined by Western blot. The loaded proteins in the two “- or +” lines were the same but from the different plates. (C) mRNA levels of downstream targets of sXBP-1 in IER3IP1 KD cells. 4 days after IER3IP1 KD in MIN6 cells, mRNA levels of downstream targets of sXBP-1, Erdj4, p58IPK, EDEM & Sec61a, were determined by qRT-PCR. Values are mean ± SEM. (D) Effect of IER3IP1 suppression on the Perk arm of the UPR. 4 days after IER3IP1 KD in MIN6 cells, phosphorylation of Perk & eIF2a, & ATF4 were determined by Western blot. The loaded proteins in the two “- or +” lines were the same but from the different plates. (E) IER3IP1 KD decreases UPR activation induced by ER stress in MIN6. 4 days after IER3IP1 KD, MIN6 cells were treated with 10 μg/ml tunicamycin (TC) or 1 μM thapsigargin (TG) for 6 hours. Then the expression levels of Chop, sXBP-1, uXBP-1 & IER3IP1 were determined by Western blot. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28915629), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - ATG5 & ATG7 influenced on autophagy, apoptosis & cell cycle through PERK/Nrf2 signaling. a The ER stress proteins were analysed by western blotting in Ad-GFP, Ad-ATG5 + Ad-ATG7,Ad-ATG5 + Ad-ATG7 + siPERK,Ad-ATG5 + Ad-ATG7 + siNrf2 & Ad-ATG5+ Ad-ATG7 + siPERK+siNrf2 induced chondrocytes. b The levels of ER stress proteins were normalized to beta -actin. c Determination of autophagy & apoptosis proteins expression by western blotting in Ad-GFP, Ad-ATG5 + Ad-ATG7, Ad-ATG5 + Ad-ATG7 + siPERK,Ad-ATG5 + Ad-ATG7 + siNrf2 & Ad-ATG5+ Ad-ATG7 + siPERK+ siNrf2 induced chondrocytes. dThe levels of related proteins were normalized to beta -actin. e FCM analysis was used to calculate the percentage of apoptotic cells at the time point of 24 h. The apoptosis rate were increased when combined treatment with silencing of PERK or Nrf2. Experiments were repeated 3 times, Representative images are shown. f Analysis of cell apoptosis. Data come from 3 independent experiments. g FCM analysis indicated that the S phase percentage were decreased compared to that of the control groups when combine infected with silencing of PERK or Nrf2 in C28I2 cells. Experiments were repeated 3 times, Representative images are shown. h Percentage of cells at each phase in different groups. *P < 0.05, **P < 0.01 compared with the controls. Values are means ± SD n = 3). (1:Ad-GFP, 2:Ad-ATG5 + Ad-ATG7, 3:Ad-ATG5 + Ad-ATG7 + siPERK, 4:Ad-ATG5 + Ad-ATG7 + siNrf2, 5:Ad-ATG5 + Ad-ATG7 + siPERK+ siNrf2) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31060556), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Thapsigargin treatment induces calpain activation & autophagy. (A) Control & shCAPNS1 cells were treated with 100 nM thapsigargin for 10, 20 & 30 min & the lysates subjected to western blot analysis to quantify the precursor & active form of CAPN1. The ratio between active CAPN1 & tubulin is reported below each lane. (B) Time course measurement of intracellular calcium concentration. The plot indicates the ratio of mean Indo Violet/Indo Blue emission values of each cell population at individual time points (minutes). The ratio corresponds to the relative calcium concentration of control & shCAPNS1 U2OS cells before & after addition of thapsigargin (100 nm final). Moving average was used as smoothing method. (C) shCAPNS1 cells were treated with 100 nM thapsigargin for 30, 60 & 120 min & the lysates subjected to western blot analysis to detect LC3, p62, pIRE, CAPNS1 & tubulin. (D) Levels of p62 & LC3 normalized to tubulin levels. Image collected & cropped by CiteAb from the following publication (//journals.biologists.com/bio/article/doi/10.1242/bio.022806/256614/Calpain-mobilizes-Atg9-Bif-1-vesicles-from-Golgi), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Abnormal Paneth cells show ER stress.(A, B) Representative transmission electron microscopy images of Paneth cells at the base of ileal crypts in (A) ICR & (B) SAMP1/YitFc mice. Scale bars indicate 2 μm. (C, D) Quantitative analysis of (C) granule number & (D) ER lumen diameter in Paneth cells (n = 3/each week for SAMP1/YitFc mice). For the measurements, three Paneth cells were randomly selected from each mouse. (E) SDS–PAGE Western blot analysis of ER stress markers, pIRE1 alpha , ATF4, cleaved-ATF6, & GRP78 in ileal crypts (n = 4/each group). Total-IRE1 alpha & HPRT1 was used as loading control. (F) Relative expression level of ER stress markers calculated from the band intensity. Error bars represent mean ± SEM. (C, D, F) Statistical significance was evaluated by t test in (C, D), & one-way ANOVA followed by Tukey’s post hoc test in (F). P < 0.05 was considered statistically significant. *P < 0.05, †P < 0.01, §P < 0.001. E, ER; G, granules; N, nucleus; n.s., not significant. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32345659), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: IRE1 alpha [p Ser724] Antibody - BSA Free [NB100-2323] - Hypoxia-induced downregulation of gamma -taxilin & alpha NAC & the subsequent ER responses are GSK-3 beta -dependent. (a) GSK-3 beta activation in hypoxia. Western blot analysis shows downregulation of phosphorylated (Ser 9) GSK-3 beta (phospho-GSK-3 beta ) in hypoxic (0–48 h) SK-N-SH & hypoxic (0–24 h) HeLa S3 cells. (b) Inhibition of GSK-3 beta activation by lithium chloride (LiCl, 100 mM) or CHIR-99021 (CHIR, 15 or 30 μM) maintained gamma -taxilin & alpha NAC protein levels in hypoxic SK-N-SH & HeLa S3 cells. (c) GSK-3 beta inhibition suppresses the expression of ER stress response proteins that are induced in hypoxic SK-N-SH cells. (d) GSK-3 beta RNA interference almost completely blocked the protein expression & restored the expression of gamma -taxilin & alpha NAC proteins in hypoxic HeLa S3 cells Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25880086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Dr, Gt, Ma, Pm, Pm, Rb, SyHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Dr, Gt, Ma, Pm, Pm, Rb, SyHa

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-2323AF350
Alexa Fluor 405	NB100-2323AF405
Alexa Fluor 488	NB100-2323AF488
Alexa Fluor 532	NB100-2323AF532
Alexa Fluor 594	NB100-2323AF594
Alexa Fluor 647	NB100-2323AF647
Alexa Fluor 700	NB100-2323AF700
Alexa Fluor 750	NB100-2323AF750
Allophycocyanin	NB100-2323APC
Biotin	NB100-2323B
CoraFluor 1	NB100-2323CL1
DyLight 350	NB100-2323UV
DyLight 405	NB100-2323V
DyLight 488	NB100-2323G
DyLight 550	NB100-2323R
DyLight 594	NB100-2323DL594
DyLight 650	NB100-2323C
DyLight 680	NB100-2323FR
DyLight 755	NB100-2323IR
FITC	NB100-2323F
HRP	NB100-2323H
Janelia Fluor 525	NB100-2323JF525
Janelia Fluor 549	NB100-2323JF549
Janelia Fluor 585	NB100-2323JF585
Janelia Fluor 635	NB100-2323JF635
Janelia Fluor 646	NB100-2323JF646
Janelia Fluor 669	NB100-2323JF669
PE	NB100-2323PE
PerCP	NB100-2323PCP
mFluor Violet 450 SE	NB100-2323MFV450
mFluor Violet 500 SE	NB100-2323MFV500
mFluor Violet 610 SE	NB100-2323MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-2323SS

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NB500-201

Survivin Antibody - BSA Free

(22 Reviews)

(374 Publications)

0.1 ml

NB500-201

<b>Biological Strategies Validation, Genetic Strategies Validation and Orthogonal Strategies Validation.</b>Knockdown Validated: Survivin Antibody [NB500-201] - Western blot analysis using Survivin and flotillin antibodies was performed on lysates of DMSO- and PTX-treated MDAMB231 cells ectopically expressing either control siRNA or Survivin siRNA (panels labeled WCL), as well as on the exosomes these cells generated (panels labeled Exos). Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2072-6694/8/12/111) licensed under a CC-BY license.

Immunohistochemistry: Survivin Antibody [NB500-201] - Protein expression pattern of Survivin in HCC tissues and non-neoplastic liver parenchyma. IAP members immunoreactivity was estimated by tissue microarray in a subset of HCC patients (n = 40). A-D, Representative Survivin cytoplasmatic immunostaining in a tumor core (A), in a tumor proximal to cirrhosis (C, N: cirrhosis, K: HCC), and in adjacent and long-distance non-neoplastic parenchyma (B and D, respectively). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/19397802) licensed under a CC-BY license.

<b>Genetic Strategies Validation. </b>Knockdown Validated: Survivin Antibody [NB500-201] - Anti-tumor effects and functional evidence of sCA-survivin-siRNA in HCT116 and HT29 solid tumor models. Immunostaining of survivin in the tumor tissues on day 19 using [NB500-201]. Scale bar, 50 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0116022) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: Survivin Antibody [NB500-201] - Analysis using the HRP conjugate of [NB500-201]. Staining of Telophase with accumulation of survivin in the midbodies of two daughter cells. Survivin detection using [NB500-201].

Western Blot: Survivin Antibody [NB500-201] - Analysis of 30ug of HeLa whole cell lysate [NB800-PC1] using rabbit polyclonal [NB500-201] at 1ug/ml. Detection was performed using ECL method with 1 minute exposure. Band detected at higher molecular weight than the predicted MW (16 kDa).

Immunohistochemistry-Paraffin: Survivin Antibody [NB500-201] - Immunohistochemical staining of Survivin in human rectal cancer using [NB500-201] and DAB with hematoxylin counterstain.

Flow Cytometry: Survivin Antibody [NB500-201] - An intracellular stain was performed on HeLa cells with [NB500-201] and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550.

Western Blot: Survivin Antibody [NB500-201] - Western blot analysis using Survivin Antibody [NB500-201]. Effects of siRNA transfection on the expression of Bcl-xL and survivin. Bcl-xL and survivin protein content detected by Western Blotting 48 h after transfection. Bcl-xL and survivin levels are shown normalized to the reference protein beta-actin and relative to the ns-si control. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/14/6/12297) licensed under a CC-BY license.

Western Blot: Survivin Antibody [NB500-201] - Analysis of Survivin in human hepatocytes from cancer patient (left) and HeLa cell lysate (right) using [NB500-201]. Image from verified customer review. Note: bands detected at higher molecular weight than predicted (16 kDa)

Western Blot: Survivin Antibody [NB500-201] - Western blot analysis using [NB500-201]. Survivin protein is degraded by ubiquitin proteasome in serum-free and serum-containing media. Western blots and protein quantitation graphs showing that addition of MG-132 proteasome inhibitor (10 uM) extended survivin protein half-life in serum-free and serum-containing media. One representative western blot out of triplicate experiments was shown for each treatment and condition. *Indicates the time at which survivin protein is half of the amount at 0 hours (half-life), P 100. ** p-value 100. ** p-value < .01 using student t-test. H. PFGE analysis of EBV episomes & linears at 7 days post-transduction with lentivirus shRNA for shControl or shSurvivin in MUTU I or RAJI cell lines. Human alpha satellite DNA is used for DNA loading control Western blot of Survivin & Actin are shown below for each cell at 7 days post-transduction. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28077791), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Survivin Antibody - BSA Free [NB500-201] - The p53-p21 axis facilitates downregulation of survivin via cell cycle(A) HCT116 wild typeand p21-deficient (p21-/-) cells were treated with 5 μM L-OHP for 24 hours. Whole cell lysates were analyzed with antibodies against p53, p21, & survivin; vinculin serves as loading control. (B) Cell cycle distribution was analyzed after 24 hours treatment by flow cytometry analysis (n = 3). (C) To induce p21, RKO p21ind cells were treated with 3 nM Muristerone A for 24 hours & tested for the levels of p21 & survivin; vinculin, loading control. (D)BIRC5 mRNA levels were analyzed by quantitative real-time PCR after 24 hours treatment with MurA in RKO p21ind cells (n = 3). (E) Cell cycle distribution was measured by flow cytometry analyses of cellular DNA content (n = 3). (F) Scheme summarizing the supposed mechanisms of survivin regulation after L-OHP & CPT-11 treatment. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29963241), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Survivin Antibody - BSA Free [NB500-201] - Apoptosis & survival signaling after L-OHP & CPT-11(A) Western blot analysis using antibodies against p53 & pro-apoptotic BAX & PIG-3 after treatment with 5 μM L-OHP or 10 μM CPT-11. (B) Immunodetection of NF-kappa B p65, RELB & anti-apoptotic survivin, XIAP, BCL-XL & MCL1; vinculin serves as loading control. (C) Effects of increasing doses L-OHP & CPT-11 on caspase-3 & PARP1 cleavage after 24 hours treatment; alpha -tubulin serves as loading control. (D) Cells were treated with a combination of L-OHP & the caspase-inhibitor Z-VAD-FMK (50 μM). Immunodetection of survivin, p53 & full-length caspase-3 was conducted. Detection of apoptosis was determined by cleavage products of caspase-3 & PARP1; beta -actin serves as loading control. Please note: Figure 4A & 4B, as well as Supplementary Figure 2A show signals acquired by different detection methods, but originate from the same Western blots. This is due to a switch in the immunoblot chemiluminescence detection system from X-ray films (darker background) to a CCD camera system (Fusion Solo S, Vilber Lourmat; lighter background). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29963241), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Survivin Antibody - BSA Free [NB500-201] - Anti-tumor effects & functional evidence of sCA-survivin-siRNA in HCT116 & HT29 solid tumor models.(A) For western blot analysis, HCT116 cells were seeded into 6-well plates & transfected with 100 pmol/well of either control or survivin siRNA by Lp (Lipofectamine 2000) or sCA. Actin blots served as loading controls. (B) For proliferation assays, cells were uniformly seeded into 96-well plates (1 × 104 cells/well), & 5 pmol/well siRNA was used. Cell viability was examined at 48 & 72 h by WST-8 assay. Data represent mean ± SEM. *P = 0.0294, **P = 0.0304 (n = 4, Wilcoxon rank test). (C) In vivo tumor growth. Each vehicle, carrying 15 μg of control siRNA or survivin siRNA, was administered by intravenous injection to mice with HCT116 tumors. Data represent the mean ± SEM (n = 10 tumors, Wilcoxon rank test). (D) Immunostaining of survivin in the tumor tissues on day 19. Scale bar, 50 μm. (E) In vivo live imaging of sCA-siRNA (6-FAM labeled) in HT29 tumor by multiphoton microscopy at 90 min. (F) Fluorescent detection of naked-siRNA (6-FAM labeled) or sCA-siRNA (6-FAM labeled) in the HT29 tumor at 4 h. Green: 6-FAM labeled siRNA; Red: microvasculature; Blue: DAPI stained nuclei. Scale bar, 50 μm. (G) Mice were administered with 40 μg of naked-survivin-siRNA or sCA-survivin-siRNA on days 0, 1, & 2. Tumors were removed on day 3, & western blot analysis for survivin was performed. (H) Many tumor cells treated with sCA-survivin-siRNA (6-FAM labeled) had condensed nuclei & were positive for TUNEL assay. Scale bar, 50 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25738937), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Survivin Antibody - BSA Free [NB500-201] - Survivin is essential for the salinomycin-induced apoptosis in UM cells. a UM cells were treated with salinomycin for 24 h & the protein levels of apoptosis-related proteins were detected by Western blot. b & c UM cells were transduced with lentiviral pTSB-Survivin cDNA (b), Survivin-shRNA constructs (c), or their corresponding empty vectors, & then incubated in the presence of puromycin (1 μg/mL) for 5 days to reach stable clones. Such survivin-manipulated cells were then exposed to salinomycin for 24 h, & subjected to trypan blue exclusion assay (left) & Western blot assay (right). Data represent mean ± SD. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001, Student’s t test. d qRT-PCR analysis of BIRC5 mRNA level was done in the 92.1 & Mel270 cells treated with salinomycin for 24 h. **, P < 0.01; ***, P < 0.001, one-way ANOVA, post hoc comparisons, Tukey’s test Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31718679), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Survivin Antibody - BSA Free [NB500-201] - Anti-tumor effects & functional evidence of sCA-survivin-siRNA in HCT116 & HT29 solid tumor models.(A) For western blot analysis, HCT116 cells were seeded into 6-well plates & transfected with 100 pmol/well of either control or survivin siRNA by Lp (Lipofectamine 2000) or sCA. Actin blots served as loading controls. (B) For proliferation assays, cells were uniformly seeded into 96-well plates (1 × 104 cells/well), & 5 pmol/well siRNA was used. Cell viability was examined at 48 & 72 h by WST-8 assay. Data represent mean ± SEM. *P = 0.0294, **P = 0.0304 (n = 4, Wilcoxon rank test). (C) In vivo tumor growth. Each vehicle, carrying 15 μg of control siRNA or survivin siRNA, was administered by intravenous injection to mice with HCT116 tumors. Data represent the mean ± SEM (n = 10 tumors, Wilcoxon rank test). (D) Immunostaining of survivin in the tumor tissues on day 19. Scale bar, 50 μm. (E) In vivo live imaging of sCA-siRNA (6-FAM labeled) in HT29 tumor by multiphoton microscopy at 90 min. (F) Fluorescent detection of naked-siRNA (6-FAM labeled) or sCA-siRNA (6-FAM labeled) in the HT29 tumor at 4 h. Green: 6-FAM labeled siRNA; Red: microvasculature; Blue: DAPI stained nuclei. Scale bar, 50 μm. (G) Mice were administered with 40 μg of naked-survivin-siRNA or sCA-survivin-siRNA on days 0, 1, & 2. Tumors were removed on day 3, & western blot analysis for survivin was performed. (H) Many tumor cells treated with sCA-survivin-siRNA (6-FAM labeled) had condensed nuclei & were positive for TUNEL assay. Scale bar, 50 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25738937), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ca, Fe, Gp, Ha

Species Glossary

Reactivity: Hu, Mu, Rt, Ca, Fe, Gp, Ha

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	NB500-201AF350
Alexa Fluor 405	NB500-201AF405
Alexa Fluor 488	NB500-201AF488
Alexa Fluor 532	NB500-201AF532
Alexa Fluor 594	NB500-201AF594
Alexa Fluor 647	NB500-201AF647
Alexa Fluor 700	NB500-201AF700
Alexa Fluor 750	NB500-201AF750
Allophycocyanin	NB500-201APC
Biotin	NB500-201B-0.1ml
DyLight 350	NB500-201UV
DyLight 405	NB500-201V-0.1ml
DyLight 488	NB500-201G-0.1ml
DyLight 550	NB500-201R-0.1ml
DyLight 594	NB500-201DL594
DyLight 650	NB500-201C-0.1ml
DyLight 680	NB500-201FR
DyLight 755	NB500-201IR
FITC	NB500-201F-0.1ml
HRP	NB500-201H-0.1ml
Janelia Fluor 549	NB500-201JF549
Janelia Fluor 646	NB500-201JF646
PE	NB500-201PE
PerCP	NB500-201PCP
mFluor Violet 450 SE	NB500-201MFV450
mFluor Violet 500 SE	NB500-201MFV500
mFluor Violet 610 SE	NB500-201MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB500-201SS

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NBP1-04676

PGC1 alpha Antibody - BSA Free

(11 Reviews)

(278 Publications)

0.1 ml

NBP1-04676

Mitochondrial content and biogenesis. Immunoblot analysis of PGC1 alpha in MEFs (n = 7; t = 5.764; df = 6). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-12662-3), licensed under a CC-BY license.

HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-PGC1 alpha Antibody NBP1-04676 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Analysis of a FFPE section of mouse prostate using rabbit polyclonal PGC1 alpha Antibody at 1:200 dilution. The antibody generated an expected strong nuclear (punctate in some cells) and a weak cytoplasmic staining in the glandular cells lining of tubule-alveolar gland.

An intracellular stain was performed on A431 cells with PGC1 alpha Antibody NBP1-04676 (blue) and matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

<b>Biological Strategies Validation. </b>Vehicle or RU 486 (5 mg/kg) injected mice were presented with/without corticosterone (10 mg/kg) for 3 days. The expressions of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha NBP1-04676) and NIX (BNIP3L NBP1-88558) were visualized via western blotting. Loading control is beta-actin. n = 5. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33473105/) licensed under a CC-BY license.

Total protein from human adipose and skeletal muscle tissue, HeLa and A431 cells lines was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL PGC1 alpha Antibody (Molecular weight: 91 KDa) in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence

Western Blot of total protein from murine skeletal muscle tissue using PGC1 alpha Antibody. Image from verified customer review.

HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with PGC1 alpha Antibody [NBP1-04676] at a 1:200 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) [NB100-690] was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Analysis of PGC1 alpha Antibody in mouse prostate using DAB with hematoxylin counterstain.

An intracellular stain was performed on HeLa cells with NBP1-04676AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

An intracellular stain was performed on HepG2 cells with PGC1 alpha Antibody and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody.

<b>Genetic Strategies Validation. </b>Western blot shows lysates of A431 human squamous carcinoma parental cell line and PGC1 alpha knockout (KO) A431 cell line. PVDF membrane was probed with 1:1000 of Rabbit Polyclonal PGC1 alpha Antibody (Catalog # NBP1-04676) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog #HAF008). Specific band was detected for PGC1 alpha at approximately 105 kDa (as indicated) in the parental A431 cell line, but is not detectable in the knockout A431 cell line. This experiment was conducted under reducing conditions.

DM1-derived fibroblasts have no changes in mitochondria biogenesis. (A, B) Representative images of immunofluorescence of TOMM20, and PGC1-alpha in DM1 and control fibroblasts (n=3). (C) Medium fluorescence intensity of MitoTracker Red FM in control (n=3) and DM1 cells (n=5) and (D) of Rhodamine 123 in DM1 and control fibroblasts (n=3). (E) mRNA levels of TFAM transcription factor (n=3). (F) mRNA levels of OPA1, MFN1, MFN2, DRP1 and PARKIN in DM1 and control fibroblasts (n≥2).

A431 human skin carcinoma cell line was stained with Rabbit anti-PGC1 alpha Affinity-purified Polyclonal Antibody conjugated to Phycoerythrin (Catalog # NBP1-04676PE, blue histogram) or matched control antibody (Catalog # NBP2-24983, orange histogram).

Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Effect of butein T on SC adipose tissue browning in the ThermoMouse. SC fat was obtained from ThermoMouse exposed for 4 days to butein (10 or 20 mg/kg; n = 4) or vehicle (n = 2). A BAT lysate of a ThermoMouse was used as a positive control for UCP-1 protein levels. Tissues were homogenized & protein lysates (50 µg for SC fat & 10 µg for BAT) were used in western blotting for PGC1-alpha (a) & UCP-1 (c). Panels (b) & (d) illustrate the densitometric quantitation of the PGC1-alpha & UCP-1 western blot, respectively, normalized for the levels of the loading control alpha -tubulin. Abbreviations: subcutaneous (SC); brown adipose tissue (BAT); peroxisome proliferator-activated receptor-gamma coactivator 1-alpha (PGC1-alpha ) & uncoupling protein-1 (UCP-1). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31094273), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Expression of the mitochondrial biogenesis proteins: SIRT1, PGC1 alpha , TFAM & SIRT3 in H1299, H1299r, P31 & P31r cell lysates as determined by immunoblot analysisWhole cell lysates were prepared from confluent cultures of H1299, H1299r, P31 & P31r cells. Proteins (90 μg) were resolved in 10% SDS-PAGE gels & transferred to a PVDF membrane. Blots were probed for SIRT1, PGC1 alpha , TFAM & SIRT3 or the loading control gamma -tubulin. Figure shows representative blots of (A) of SIRT1, (B) PGC1 alpha , (C) SIRT3, (D) TFAM protein expression; each from three independent experiments. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.21885), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Maternal exercise during pregnancy on mitochondrial biogenesis in the fetal hearts. (A) Levels of relative mRNA expression measured by qRT‐PCR. n = 9–12/group. Maternal exercise during pregnancy did not alter levels of mRNA in Ppargc1a & Tfam, while it significantly upregulated the levels of mRNA in Nrf1 & Nrf2. (B–D) Densitometric analyses of protein expression levels relative to the sedentary group with representative images of western blots were shown. No significant differences in PGC‐1 alpha , NRF1, & NRF2 (P > 0.05). n = 5–6/group. * P < 0.05, significantly different from the sedentary group. Black bar: fetal hearts from sedentary dams; gray bar: fetal hearts from exercised dams. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28292876), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: PGC1 alpha Antibody - BSA Free [NBP1-04676] - DM1-derived fibroblasts have no changes in mitochondria biogenesis. (A, B) Representative images of immunofluorescence of TOMM20, & PGC1-alpha in DM1 & control fibroblasts (n=3). (C) Medium fluorescence intensity of MitoTracker Red FM in control (n=3) & DM1 cells (n=5) & (D) of Rhodamine 123 in DM1 & control fibroblasts (n=3). (E) mRNA levels of TFAM transcription factor (n=3). (F) mRNA levels of OPA1, MFN1, MFN2, DRP1 & PARKIN in DM1 & control fibroblasts (n≥2). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32310829), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Effect of BGP-15 treatment on the regulation of mitochondrial biogenesis in NRCM cells. Western blot analysis of PGC-1-alpha , CREB, & VDAC proteins as well as densitometric evaluation is shown. GAPDH was used as a loading control. Control group: cells without any treatment; BGP-15 group: cells with only 50 μM BGP-15 for 0.5 hours; H2O2 group: cells with 150 μM H2O2 for 0.5 hours; H2O2+BGP-15 group: cells with 150 μM H2O2 & 50 μM BGP-15 for 0.5 hours. Values are mean ± SEM (n = 4). ∗p < 0.05 vs. Control, ∗∗p < 0.01 vs. Control, §§p < 0.01 vs H2O2 group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33728024), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Feeding the diabetes mellitus (DM) mice with the RSV‐enriched diet improved mitochondrial biogenesis in the quadriceps muscle. A) Representative western blots using antibodies against nuclear respiratory factor (NRF‐1), ATP synthase, H+ transporting, mitochondrial F1 complex, beta polypeptide (ATP5B), cytochrome c oxidase subunit IV (Cox IV), peroxisome proliferator‐activated receptor‐ gamma coactivator 1‐ alpha (PGC‐1 alpha ), & GAPDH. After quantification, the B) NRF‐1/GAPDH, C) ATP5B/GAPDH, D) Cox IV/GAPDH, & E) PGC‐1 alpha /GAPDH ratios were calculated in mice fed either the control (CTL) or RSV‐enriched diet. GAPDH was used as a loading control. F) Representative western blots using antibodies against mitochondrial transcription factor A (mtTFA) & voltage‐dependent anion channel (VDAC), & the mtTFA/VDAC ratio was determined. The mitochondrial protein VDAC was used as a loading control. Data are expressed as fold change versus CTL & reported as mean ± SD, n = 4–6 per group; *p < 0.05; **p < 0.01 between CTL & DM groups. #p < 0.05 between DM & DM+RSV groups; ††p < 0.05 between CTL+RSV & DM+RSV groups. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29578301), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Effects of SRTAW04 on expression of markers of mitochondrial & anti-oxidant function. (a) Western blot of protein extracts from optic nerve & retina of control (lanes 1–4), MHV-A59 infected (lanes 5–8), & MHV-A59 infected + SRTAW04-treated (lanes 9–12) mice. Average levels of SDHb measured by Western blotting (n = 4/group) showed a significant (*p < 0.05) decrease in protein extracts from optic nerves (a,b) & retinas (a,d) of MHV-A59 infected mice 7 days post-inoculation, compared to control mice. MHV-A59 infected mice treated with SRTAW04 (100 mg/kg/day) showed a significant increase (@p < 0.05) of SDHb protein expression compared to untreated MHV-A59 infected mice. There is a significant decrease (*p < 0.05) in expression of SOD2 (n = 4/group) in optic nerves (a,c) & retinas (a,e) of MHV-A59 infected mice compared to control mice, & treatment with SRTAW04 significantly (@p < 0.05) attenuates that change. PGC1-alpha expression shows a significant (*p < 0.05) decrease in retinas (a,f) (n = 4/group) during MHV-A59 infection & treatment with SRTAW04 for 7 days significantly (@p < 0.05) increases the PGC1-alpha protein levels. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24383546), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - RSV treatment increases markers of mitochondrial function in stressed RGC-5 cells. Staurosporine-differentiated RGC-5 cells were cultured with or without 500 μM H2O2, & with or without 0.25 μM RSV for 24 h. (A) Western blot analysis shows a significant decrease in SDH expression (**p < 0.01) during H2O2 treatment which is attenuated by treatment with RSV (*p < 0.05). (B) Western blot analysis shows similar effects on SOD2 expression. The significant decrease (**p < 0.01) during H2O2 treatment, compared to controls, is not found in cells treated with RSV (*p < 0.05). (C) Protein extracts were immunoprecipitated with anti-PGC-1 alpha antibodies, blotted, & hybridized with anti-PGC-1 alpha & anti-acetylated lysine antibodies to assess the acetylation state of PGC-1 alpha . H2O2 treatment significantly increases the proportion of acetylated PGC-1 alpha (*p < 0.05) compared to controls, & RSV & SRTAW04 treatment each prevent this acetylation (*p < 0.05). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23293585), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Pequi extract increases expression of antioxidant enzymes in HCAEC. Cells were cultured, pre-treated with pequi extract (25 μg/mL) for 24 h under normoxia conditions, & total protein content was extracted for Western blots analysis for Superoxide dismutase (SOD)-1 (A), SOD-2 (B), catalase (C), glutathione peroxidase—GPx (D), sirtuin 1—SIRT-1 (E), peroxisome-proliferator-activated receptor gamma coactivator 1-alpha (PCG1-alpha ) (F), glycogen synthase kinase 3 beta (GSK3 beta ) (G), & microtubule-associated protein 1A/1B-light chain 3 (LC3) (H). Western blots were normalized using alpha -tubulin or glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as loading control. Five independent Western blot experiments were carried for each condition. Representative images are shown here. Statistical analysis was performed using Student’s t-test (p < 0.05). OD: Optical Density. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35326129), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Immunocytochemistry/ Immunofluorescence: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Role of PGC1 alpha in NIX-dependent mitophagy.a–e Nontargeting (NT) or GR siRNA was transfected to hippocampal neurons & SH-SY5Y cells for 24 h prior to corticosterone & cortisol for 12 h, respectively. a, b Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha ) expression was detected in western blot where beta -actin was used as a loading control in both cell types. n = 5. c Colocalization of PGC1 alpha (red) & DAPI (blue) in hippocampal neurons was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. d Colocalization of PGC1 alpha (green) & DAPI (blue) in SH-SY5Y was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. e PGC1 alpha protein expressions in subcellular fraction samples were detected by western blotting. Lamin A/C & alpha -tubulin were used as a nuclear & cytosolic loading control, respectively. n = 5. f, g SH-SY5Y cells were transfected with pcDNA3.1/c-eGFP or pcDNA3.1/PPARGC1A-c-eGFP vector for 24 h prior to cortisol treatment for 24 h. f NIX expression was detected in western blot where beta -actin was used as a loading control. n = 5. g TOMM20 levels were detected by western blot. Loading control for western blot is beta -actin. n = 5. All blots & immunofluorescence images are representative. n = 5 from independent experiments with two technical replicates each. Quantitative data are presented as a mean ± S.E.M. Two-sided two-way ANOVA was conducted. ** indicates p < 0.01 versus control. #, ## indicates p < 0.05, p < 0.01 versus corticosterone in hippocampal neurons & cortisol in SH-SY5Y, respectively. Data are provided as a Source data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33473105), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Role of PGC1 alpha in NIX-dependent mitophagy.a–e Nontargeting (NT) or GR siRNA was transfected to hippocampal neurons & SH-SY5Y cells for 24 h prior to corticosterone & cortisol for 12 h, respectively. a, b Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha ) expression was detected in western blot where beta -actin was used as a loading control in both cell types. n = 5. c Colocalization of PGC1 alpha (red) & DAPI (blue) in hippocampal neurons was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. d Colocalization of PGC1 alpha (green) & DAPI (blue) in SH-SY5Y was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. e PGC1 alpha protein expressions in subcellular fraction samples were detected by western blotting. Lamin A/C & alpha -tubulin were used as a nuclear & cytosolic loading control, respectively. n = 5. f, g SH-SY5Y cells were transfected with pcDNA3.1/c-eGFP or pcDNA3.1/PPARGC1A-c-eGFP vector for 24 h prior to cortisol treatment for 24 h. f NIX expression was detected in western blot where beta -actin was used as a loading control. n = 5. g TOMM20 levels were detected by western blot. Loading control for western blot is beta -actin. n = 5. All blots & immunofluorescence images are representative. n = 5 from independent experiments with two technical replicates each. Quantitative data are presented as a mean ± S.E.M. Two-sided two-way ANOVA was conducted. ** indicates p < 0.01 versus control. #, ## indicates p < 0.05, p < 0.01 versus corticosterone in hippocampal neurons & cortisol in SH-SY5Y, respectively. Data are provided as a Source data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33473105), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - EMPA treatment increased mitochondrial biogenesis. (A) Left, Transmission electron microscopy showing the morphology of mitochondria in sham, sham + EMPA, TAC & TAC + EMPA groups & Right, quantitative analysis of mitochondrial size & counts. Results are expressed as mean ± SEM, n = 3–5, *p < 0.05 vs. corresponding sham, †p < 0.05 vs. corresponding TAC. (B) Left, Representative blots of mitochondrial biogenesis-related proteins & Right, quantitative results. (C) Relative mRNA levels of PGC1 alpha , NRF1, TFAM & COX1. Results are expressed as mean ± SEM, n = 5–7, *p < 0.05 vs. corresponding sham group, †p < 0.05 vs. corresponding TAC vehicle group. One-way ANOVA & Tukey post hoc test. EMPA, empagliflozin; SEM, standard error of the mean; TAC, transverse aortic constriction; PGC1-alpha , peroxisome proliferator-activated receptor gamma coactivator 1-alpha; NRF-1, nuclear respiratory factor 1; TFAM, mitochondrial transcription factor A; COX1, cyclooxygenase1; GAPDH, glyceraldehyde 3-phosphate dehydrogenase. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35647080), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Immunocytochemistry/ Immunofluorescence: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Role of PGC1 alpha in NIX-dependent mitophagy.a–e Nontargeting (NT) or GR siRNA was transfected to hippocampal neurons & SH-SY5Y cells for 24 h prior to corticosterone & cortisol for 12 h, respectively. a, b Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha ) expression was detected in western blot where beta -actin was used as a loading control in both cell types. n = 5. c Colocalization of PGC1 alpha (red) & DAPI (blue) in hippocampal neurons was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. d Colocalization of PGC1 alpha (green) & DAPI (blue) in SH-SY5Y was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. e PGC1 alpha protein expressions in subcellular fraction samples were detected by western blotting. Lamin A/C & alpha -tubulin were used as a nuclear & cytosolic loading control, respectively. n = 5. f, g SH-SY5Y cells were transfected with pcDNA3.1/c-eGFP or pcDNA3.1/PPARGC1A-c-eGFP vector for 24 h prior to cortisol treatment for 24 h. f NIX expression was detected in western blot where beta -actin was used as a loading control. n = 5. g TOMM20 levels were detected by western blot. Loading control for western blot is beta -actin. n = 5. All blots & immunofluorescence images are representative. n = 5 from independent experiments with two technical replicates each. Quantitative data are presented as a mean ± S.E.M. Two-sided two-way ANOVA was conducted. ** indicates p < 0.01 versus control. #, ## indicates p < 0.05, p < 0.01 versus corticosterone in hippocampal neurons & cortisol in SH-SY5Y, respectively. Data are provided as a Source data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33473105), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Role of PGC1 alpha in NIX-dependent mitophagy.a–e Nontargeting (NT) or GR siRNA was transfected to hippocampal neurons & SH-SY5Y cells for 24 h prior to corticosterone & cortisol for 12 h, respectively. a, b Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha ) expression was detected in western blot where beta -actin was used as a loading control in both cell types. n = 5. c Colocalization of PGC1 alpha (red) & DAPI (blue) in hippocampal neurons was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. d Colocalization of PGC1 alpha (green) & DAPI (blue) in SH-SY5Y was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. e PGC1 alpha protein expressions in subcellular fraction samples were detected by western blotting. Lamin A/C & alpha -tubulin were used as a nuclear & cytosolic loading control, respectively. n = 5. f, g SH-SY5Y cells were transfected with pcDNA3.1/c-eGFP or pcDNA3.1/PPARGC1A-c-eGFP vector for 24 h prior to cortisol treatment for 24 h. f NIX expression was detected in western blot where beta -actin was used as a loading control. n = 5. g TOMM20 levels were detected by western blot. Loading control for western blot is beta -actin. n = 5. All blots & immunofluorescence images are representative. n = 5 from independent experiments with two technical replicates each. Quantitative data are presented as a mean ± S.E.M. Two-sided two-way ANOVA was conducted. ** indicates p < 0.01 versus control. #, ## indicates p < 0.05, p < 0.01 versus corticosterone in hippocampal neurons & cortisol in SH-SY5Y, respectively. Data are provided as a Source data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33473105), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Mitochondrial content & PGC‐1 alpha levels are increased in cells with greater TFEB protein expression. (a) Mitochondrial content was measured in SH‐SY5Y cells & TFEB‐DDK cells under basal conditions by assaying citrate synthase (CS) activity. **p < 0.01 versus SH‐SY5Y; n = 6. (b) Western blotting for the mitochondrial proteins prohibitin 1 & cytochrome oxidase (COX) subunit IV also indicated that mitochondrial content was increased in TFEB‐DDK cells. (c) PGC‐1 alpha mRNA levels were measured in SH‐SY5Y cells & TFEB‐DDK cells under basal conditions by qPCR. Data normalised against beta ‐actin mRNA levels. **p < 0.01 versus SH‐SY5Y cells; n = 3. (d) SH‐SY5Y cells were treated with scrambled (scram) or TFEB siRNA for 72 h & PGC‐1 alpha mRNA levels measured. *p < 0.05 versus scram; n = 4. (e) PGC‐1 alpha protein levels were detected by western blotting in total cell lysates of SH‐SY5Y cells, TFEB‐DDK cells & a SH‐SY5Y cell line over‐expressing human PGC‐1 alpha . The fold increase in PGC‐1 alpha protein density for this blot & PGC‐1 alpha mRNA levels of the respective cell lines are reported underneath the blot. (f) TFEB‐DDK cells were treated with 10 μM carbonyl cyanide m‐chlorophenylhydrazone (CCCP) for 18 h & cytosolic & nuclear fractions prepared. PGC‐1 alpha protein was detected in cytosolic (3.5% of total volume) & nuclear fractions (35% of total volume) by western blotting. The purity of the nuclear & cytosolic fractions was assessed by western blotting using lamin A & beta ‐actin antibodies respectively. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26509433), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - PGC1 alpha & HNF4 alpha interacts & binds to Vnn1 promoter in IUGR asthmatic mice. Asthma was induced with OVA in IUGR mice. PBS induction was used as the control. Nuclear protein was extracted from lung tissues. (A) Immunoblot assay was performed for expressions of PGC1 alpha & HNF4 alpha . (B) IP was performed using mouse anti-HNF4 alpha antibody & Protein G-coupled agarose beads, followed by immunoblot with rabbit-anti-PGC1 alpha antibody. The normal mouse IgG was used as the IP control. (C) In primary cultured bronchial epithelia cells isolated from IUGR mice injected with OVA or PBS, ChIP assay was performed using mouse anti-HNF4 alpha or rabbit anti-PGC1 alpha antibodies. The normal mouse or rabbit IgG was used as the control. Graphics show the percentage of total DNA immunoprecipitated by each indicated antibody. Data are shown as mean±s.d. n=4. *P<0.01 OVA versus PBS. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32139393), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: PGC1 alpha Antibody - BSA Free [NBP1-04676] - Role of PGC1 alpha in NIX-dependent mitophagy.a–e Nontargeting (NT) or GR siRNA was transfected to hippocampal neurons & SH-SY5Y cells for 24 h prior to corticosterone & cortisol for 12 h, respectively. a, b Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha ) expression was detected in western blot where beta -actin was used as a loading control in both cell types. n = 5. c Colocalization of PGC1 alpha (red) & DAPI (blue) in hippocampal neurons was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. d Colocalization of PGC1 alpha (green) & DAPI (blue) in SH-SY5Y was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. e PGC1 alpha protein expressions in subcellular fraction samples were detected by western blotting. Lamin A/C & alpha -tubulin were used as a nuclear & cytosolic loading control, respectively. n = 5. f, g SH-SY5Y cells were transfected with pcDNA3.1/c-eGFP or pcDNA3.1/PPARGC1A-c-eGFP vector for 24 h prior to cortisol treatment for 24 h. f NIX expression was detected in western blot where beta -actin was used as a loading control. n = 5. g TOMM20 levels were detected by western blot. Loading control for western blot is beta -actin. n = 5. All blots & immunofluorescence images are representative. n = 5 from independent experiments with two technical replicates each. Quantitative data are presented as a mean ± S.E.M. Two-sided two-way ANOVA was conducted. ** indicates p < 0.01 versus control. #, ## indicates p < 0.05, p < 0.01 versus corticosterone in hippocampal neurons & cortisol in SH-SY5Y, respectively. Data are provided as a Source data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33473105), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt, Po, Gt, Ha, Sh, Sq, Ca, Eq, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Gt, Ha, Sh, Sq, Ca, Eq, Pm

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-04676AF350
Alexa Fluor 405	NBP1-04676AF405
Alexa Fluor 488	NBP1-04676AF488
Alexa Fluor 532	NBP1-04676AF532
Alexa Fluor 594	NBP1-04676AF594
Alexa Fluor 647	NBP1-04676AF647
Alexa Fluor 700	NBP1-04676AF700
Alexa Fluor 750	NBP1-04676AF750
Allophycocyanin	NBP1-04676APC
Biotin	NBP1-04676B
CoraFluor 1	NBP1-04676CL1
DyLight 350	NBP1-04676UV
DyLight 405	NBP1-04676V
DyLight 488	NBP1-04676G
DyLight 550	NBP1-04676R
DyLight 594	NBP1-04676DL594
DyLight 650	NBP1-04676C
DyLight 680	NBP1-04676FR
DyLight 755	NBP1-04676IR
FITC	NBP1-04676F
HRP	NBP1-04676H
Janelia Fluor 525	NBP1-04676JF525
Janelia Fluor 549	NBP1-04676JF549
Janelia Fluor 585	NBP1-04676JF585
Janelia Fluor 635	NBP1-04676JF635
Janelia Fluor 646	NBP1-04676JF646
Janelia Fluor 669	NBP1-04676JF669
PE	NBP1-04676PE
PerCP	NBP1-04676PCP
mFluor Violet 450 SE	NBP1-04676MFV450
mFluor Violet 500 SE	NBP1-04676MFV500
mFluor Violet 610 SE	NBP1-04676MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP1-04676SS

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NB100-2331

LC3A Antibody - BSA Free

(22 Reviews)

(306 Publications)

0.1 ml

NB100-2331

Immunohistochemistry: LC3A Antibody - BSA Free [NB100-2331] - LC3A Antibody [NB100-2331] - Staining in mouse meniscus and cartilage. Image from verified customer review.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - Autophagy mobilized during differentiation of immortalized human mesenchymal stem cells (ihMSCs). Cultured ihMSCs were stimulated to undergo osteogenesis to form osteoblasts for 30 days at 21% and 4% oxygen. LC3I and LC3II levels were assessed via immunoblot every 10 days and normalized to beta-actin to derive average LC3II band densities. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25523618/) licensed under a CC-BY license.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - LC3A Antibody [NB100-2331] - Autophagy mobilized during differentiation of immortalized human mesenchymal stem cells (ihMSCs). Cultured ihMSCs were stimulated to undergo osteogenesis to form osteoblasts for 30 days at 21% and 4% oxygen. LC3I and LC3II levels were assessed via immunoblot every 10 days and normalized to beta-actin to derive average LC3II band densities. Image collected and cropped by CiteAb from the following publication (//stemcellres.com/content/5/6/140), licensed under a CC-BY license.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - LC3A Antibody [NB100-2331] - This LC3A antibody Image shows Analysis in human cell lysates.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - LC3A Antibody [NB100-2331] - This LC3A antibody Image shows analysis of heart tissue lysates from mice which were subjected or not to 48 hours of starvation. The signal was developed using ECL method and this LC3 antibody was found to detect both forms of LC3, i.e. LC3A and LC3B. As expected, the levels of LC3B form were higher in the heart tissue lysates from starved mice.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - LC3A Antibody [NB100-2331] - Detection of HRP conjugated autophagic LC3 in mouse ES cell lysate. The atg5-/- lane (ES cells, cultured to form embryonic bodies, that are deficient in conversion of LC3-1 to LC3-11) demonstrates the specificity of NB 100-2331, as there is no detection of LC3-11. Photo courtesy of Dr. Beth Levine, UT Southwestern Medical Center.

Immunocytochemistry/Immunofluorescence: LC3A Antibody - BSA Free [NB100-2331] - LC3A Antibody [NB100-2331] - This LC3A antibody Image shows an analysis in HeLa cells using anti-LC3 antibody (red). Nuclei were counterstained with DAPI (blue).

Immunocytochemistry/Immunofluorescence: LC3A Antibody - BSA Free [NB100-2331] - LC3A Antibody [NB100-2331] - Left panel shows untreated HeLa cells. Right panel shows HeLa cells that were treated with 50 uM CQ overnight. Cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. The cells were incubated with anti-LC3A antibody at 5 ug/mL overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry: LC3A Antibody - BSA Free [NB100-2331] - LC3A Antibody [NB100-2331] - Rapamycin increases autophagy in brains of PDAPP mice. Representative epifluorescent (c200x) image of hippocampal CA1 in control- and rapamycin-fed transgenic PDAPP mice stained with an anti-LC3 antibody. An increase in LC3-immunoreactive puncta was observed in CA1 projections of transgenic PDAPP mice following rapamycin administration. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0009979) licensed under a CC-BY license.

Immunohistochemistry: LC3A Antibody - BSA Free [NB100-2331] - LC3A Antibody [NB100-2331] - Analysis in mouse renal tissue. Image from verifed customer review.

Simple Western: LC3A Antibody - BSA Free [NB100-2331] - LC3A Antibody [NB100-2331] - Image shows a specific band for LC3 in 0.5 mg/mL of Neuro2A lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - Chronic, third window RIC increases the expression of autophagosome proteins, LC3I/II & Atg5.(A) Western blots for autophagy related signaling proteins. (B) Quantification of the protein fold change in 3W RIC compared to 3W controls. Values are means ± S.E.M. n = 6–8 per group. An (*) denotes a statistically significant difference (P10 larvae/sample). Lc3 c & e, or p62 & Optineurin d protein levels were detected in absence or presence of BafA1, c & d, or in the presence or absence of Mm e. Actin was used as a loading control. Western Blots were repeated three, c & d, or two e times with protein extracts derived from independent experiments. The Lc3II/Actin or p62/Actin & Optineurin/Actin ratio, normalized to the control sample, is indicated below the blots. f–g Representative confocal micrographs & quantification of GFP-Lc3 co-localization with Mm clusters in infected dram1∆19n/∆19n & dram1+/+ larvae. The top images f show the entire region of imaging, while the bottom images f′ & f″ show details of GFP-Lc3 colocalization of Mm clusters in dram1∆19n/∆19n & dram1+/+ larvae. The arrowheads indicate GFP-Lc3-positive Mm clusters. The data is accumulated from two independent experiments (≥15 larvae/group) & represented by scatter & boxplots as detailed in the “Methods” section. Scale bars, 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32332700), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - Optn or p62 deficiency affects autophagosome formation.(A) Workflow of the experiments shown in (B-G). Larvae were treated with 100 nM of Baf A1 for 12 h from 3.5 dpf. The GPF-Lc3 negative larvae were selected to assay autophagy activity by Western blot, the GFP-Lc3 positive larvae were collected to monitor autophagic activity using confocal imaging. The red square indicates the region for confocal imaging. (B) Level of basal autophagy in WT & mutant embryos in absence or presence of Baf A1. Protein samples were extracted from 4 dpf WT & mutant larvae (>10 embryos/sample). The blots were probed with antibodies against Lc3 & Actin as a loading control. Western blots were repeated at least three times with independent extracts. (C) Quantification of Lc3-II fold changes in WT & mutant embryos in absence or presence of Baf A1. Western blot band intensities were quantified by Lab Image. Data is combined from three independent experiments. (D) Representative confocal micrographs of GFP-Lc3 puncta present in the tail fin of optn+/+, optn delta 5n/ delta 5n, p62+/+ & p62 delta 37n/ delta 37n at 4 dpf. Scale bars, 10 μm. (E). Quantification of the number of GFP-Lc3 puncta in optn+/+, optn delta 5n/ delta 5n, p62+/+ & p62 delta 37n/ delta 37n larvae with & without Baf A1 treatment. Each larva was imaged at a pre-defined region of the tail fin (as indicated by the red boxed area in Fig3 A) (≥11 larvae/group). Results are accumulated from two independent experiments. ns, non-significant, *p10 larvae/sample). The blots were probed with antibodies against Lc3 & Actin as a loading control & Lc3-II/Lc3-I ratios are indicated below. Western blots were repeated twice with independent extracts. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30818338), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - UBC9 is degraded by autophagy in epithelial cells.(A) Immunolocalization of UBC9 in ultrathin sections of HKs transduced with empty or HPV16 E6/E7 vectors. Original view (left) & higher magnification (right) of boxed regions. Gold particles are selectively enriched in autophagic structures highlighted by arrows. Scale bar = 1 μm. (B) Top: Representative WB of HaCaT cells treated with the indicated autophagic activators (left) & inhibitors (right). Activation of autophagy was monitored by the conversion of LC3 (LC3-I) to the lipidated LC3 (LC3-II) form, a marker of autophagosome production induced by autophagic stimuli [41]. LC3-II accumulation was used to verify autophagic impairment [41]. Bottom: normalized UBC9 expression. Data are expressed as fold over untreated cells. Bars represent means ± SEM of n = 4 different biological replicates. ns: not significant (Kruskal–Wallis one-way ANOVA with Dunn’s post hoc test) compared to vehicle control groups. (C) Representative WB analysis of U-2 OS or MCF7 cells treated with chloroquine. n = 3 different biological replicates. (D) Representative WB analysis of MCF7 cells transduced with scramble or ATG5 shRNA. The observed ATG5 band represents the ATG5-ATG12 conjugated form. LC3-I accumulation is reported to evidence autophagic deficiencies promoted by shATG5. n = 3 replicates of a single transduction. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1006262), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - Dram1 is required for GFP-Lc3 targeting to Mm clusters.a, b Representative confocal micrographs & quantification of GFP-Lc3 puncta in dram1∆19n/∆19n & dram1+/+ larvae in an unstimulated situation (basal autophagy, a) & following BafA1 treatment b. Each larva was imaged at a pre-defined region of the tail fin (≥11 larvae/group). Results are accumulated from two independent experiments & represented by scatter & boxplots as detailed in the “Methods” section. ns non-significant, *p 10 larvae/sample). Lc3 c & e, or p62 & Optineurin d protein levels were detected in absence or presence of BafA1, c & d, or in the presence or absence of Mm e. Actin was used as a loading control. Western Blots were repeated three, c & d, or two e times with protein extracts derived from independent experiments. The Lc3II/Actin or p62/Actin & Optineurin/Actin ratio, normalized to the control sample, is indicated below the blots. f–g Representative confocal micrographs & quantification of GFP-Lc3 co-localization with Mm clusters in infected dram1∆19n/∆19n & dram1+/+ larvae. The top images f show the entire region of imaging, while the bottom images f′ & f″ show details of GFP-Lc3 colocalization of Mm clusters in dram1∆19n/∆19n & dram1+/+ larvae. The arrowheads indicate GFP-Lc3-positive Mm clusters. The data is accumulated from two independent experiments (≥15 larvae/group) & represented by scatter & boxplots as detailed in the “Methods” section. Scale bars, 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32332700), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - Knockdown of FUT1 is associated with an increase in autophagic flux. (a) Immunoblot analysis of LC3-II & p62 levels in control & FUT1 knockdown cells. Total cell lysates from MCF-7 & T47D cells transfected with control or FUT1 siRNAs were collected at 120 & 96 h post-transfection, respectively. Equal amounts of cell lysates were then loaded in each lane & separated by SDS-PAGE. Immunoblot analysis was performed with LC3 & p62 antibodies. Actin was used as a loading control. The intensity of LC3-II & p62 protein bands on immunoblot were quantified & normalized to actin, & the relative levels of protein expression were expressed as fold change by setting the control group value to 1. Values shown are mean±S.E.M. of three independent experiments (***P100 cells per group (*P<0.05) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27560716), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - Effect of MIR376A overexpression on autophagy.(A) MCF-7 cells were co-transfected with either MIR-CNT (control plasmid) or MIR376A & GFP-LC3 plasmid, & GFP-LC3 dot formation was analyzed. White arrows indicate clusters of the GFP-LC3 dots in cells. (B) Quantification of the experiments in A. MIR376A overexpression, but not MIR-CNT expression, blocked starvation (STV)-induced autophagy (mean ± S.D. of independent experiments, n = 4, ***p<0,01). NON STV, non-starved (C) Overexpression of MIR376A resulted in a decrease in the autophagic activity of MCF-7 cells. Starvation-induced conversion of LC3-I to LC3-II in MCF-7 cells was analyzed. Tests were performed in the presence or absence of E64d (10 µg/ml) & Pepstatin A (10 µg/ml) (E+P). LC3-II/LC3-I densitometric ratios were marked. ACTB was used as a loading control. (D) MIR376A blocked starvation induced SQSTM1 degradation in MCF-7 cells. ACTB was used as a loading control. SQSTM1/ACTIN densitometric ratios were marked. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24358205), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LC3A Antibody - BSA Free [NB100-2331] - Tau clearance in vivo & autophagy function. Clearance kinetics of photoconverted Dendra-tau measured in neurons of WT-tau & AT152T-tau fish. Measurement of intensity of red Dendra-tau signal over time reflects the clearance or degradation of tau protein. (C–F) WBs for LC3-II, a well-characterized marker of autophagosome number, demonstrate that there no differences in levels of this protein between WT-tau & A152T-tau fish either at 24 hpf (pre-phenotype; C & D) or 72 hpf (post-phenotype; E & F). (E & F) Measurements of LC3-II levels in presence or absence of ammonium chloride provides a method for measuring autophagic flux. No differences observed between the two transgenic lines at 3 dpf, suggesting that autophagy functions normally in both WT-tau & A152T-tau fish (graph represents mean ± SD of four independent clutches per group for E & F & 3for C & D; two-tailed t-test). Image collected & cropped by CiteAb from the following publication (//academic.oup.com/brain/article/140/4/1128/2980948), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Am, Ca, Fi, Pl, Ze, Bv

Species Glossary

Reactivity: Hu, Mu, Rt, Am, Ca, Fi, Pl, Ze, Bv

Applications:

, , , , , , , , , , ,

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

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-2331AF350
Alexa Fluor 405	NB100-2331AF405
Alexa Fluor 488	NB100-2331AF488
Alexa Fluor 532	NB100-2331AF532
Alexa Fluor 594	NB100-2331AF594
Alexa Fluor 647	NB100-2331AF647
Alexa Fluor 700	NB100-2331AF700
Alexa Fluor 750	NB100-2331AF750
Allophycocyanin	NB100-2331APC
Biotin	NB100-2331B
CoraFluor 1	NB100-2331CL1
DyLight 350	NB100-2331UV
DyLight 405	NB100-2331V
DyLight 488	NB100-2331G
DyLight 550	NB100-2331R
DyLight 594	NB100-2331DL594
DyLight 650	NB100-2331C
DyLight 680	NB100-2331FR
DyLight 755	NB100-2331IR
FITC	NB100-2331F
HRP	NB100-2331H
Janelia Fluor 525	NB100-2331JF525
Janelia Fluor 549	NB100-2331JF549
Janelia Fluor 585	NB100-2331JF585
Janelia Fluor 635	NB100-2331JF635
Janelia Fluor 646	NB100-2331JF646
Janelia Fluor 669	NB100-2331JF669
PE	NB100-2331PE
PerCP	NB100-2331PCP
mFluor Violet 450 SE	NB100-2331MFV450
mFluor Violet 500 SE	NB100-2331MFV500
mFluor Violet 610 SE	NB100-2331MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-2331SS

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NB600-408

Collagen I Antibody

(16 Reviews)

(322 Publications)

0.1 mg

NB600-408

Rat colon tissue stained with Collagen I antibody (red) and Hoechst (blue). IHC-P image submitted by a verified customer review.

<b>Biological Strategies Validation. </b>Myofibroblast differentiation requires CD271/NGF activation. Representative alphaSMA (red) and COLLAGEN 1 (red) immunostaining with DAPI (blue) in non-treated (control), TGFbeta-treated cells with or without gamma-secretase inhibitor. Scale bars: 50 um Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-019-09992-3) licensed under a CC-BY license.

Lane 1: Human Collagen Type 1. Lane 2: None. Load: 50 ng per lane. Primary antibody: Collagen Type I antibody at 1:1000 overnight at 4C. Secondary antibody: DyLight 649 rabbit secondary antibody at 1:20000 for 30 min at RT. Block: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 139 & 130 kDa, 139 & 130 kDa for Collagen Type I. Other Band(s): Collagen Type I splice variants and isoforms.

Representative Western blot image for Collagen I, Collagen 2 and GAPDH. N/group = 8. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1111/jcmm.12367) licensed under a CC-BY license.

Macro- and micro-architecture of human adipose tissue lobule. Representative immunostaining of the human AT lobule with Collagen I (COL1), CD34, and DAPI. The position of the septa is underlined, scale bar: 100 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-019-09992-3) licensed under a CC-BY license.

Primary human cardiac fibroblast cells were stained with anti-Collagen I antibody. Cells were cultured for 3 days in DMEM with 10% fetal calf serum. ICC/IF image submitted by a verified customer review.

Cell lysate from human trabecular meshwork. Antibody at 1:1.000. WB image submitted by a verified customer review.

Patient-derived CAF and NF CDMs are architecturally and functionally distinct. (e) Collagen I (red) and fibronectin (blue) staining of patient #1 and #3 NF and CAF CDM. Scale bar, 20 μm. Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms12237) licensed under a CC-BY license.

Staining of human HaCaT-ras A5-RT3 (SCC) frozen tumor sections. Image provided by Dr. Wa'el Al Rawashdeh of RWTH Aachen University.

Livers were harvested at 9 wpi, formalin-fixed, and then cryosectioned and stained for markers of inflammation and liver fibrosis. Maximum intensity projections of 12-17 um thick liver sections with immunofluorescently labeled nuclei (DAPI white) alpha-smooth muscle actin NBP2-34760APC (yellow), CD45 (red) and Collagen I NB600-408 (top) or Collagen III/COL3A1 NB600-594 (bottom) (blue) in the liver of uninfected or 9 wpi WT mice. Scale bars represent 50 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32973293/) licensed under a CC-BY license.

Analysis in porcine burn wound lysate using anti-Collagen I alpha 1 antibody. WB image submitted by a verified customer review.

Detection of collagen I in Wistar rat hepatic stellate cells (HSC) in control (GFP-transduced) (left lane) and PPARg-transduced cell lysates (right lane). Protein staining shown below each blot depicts equal protein loading. An equal amount of the whole cell protein (100 ug) was separated by SDS-PAGE and electroblotted to nitro-cellulose membranes. Proteins were detected by incubating the membrane with anti-Collagen I antibody at a concentration of 0.2-2 ug/10 mL in TBS (100 mM Tris-HCl, 0.15 M NaCl, pH 7.4) with 5% non-fat milk. Detection occurred by incubation with a HRP conjugated secondary antibody at 1 ug/10 mL. Proteins were detected by a chemiluminescent method using the PIERCE ECL kit (Amersham Biosciences).

Balb/3T3 mouse embryonic fibroblast cell line stained with Collagen I antibody. IHC-P image submitted by a verified customer review.

Mouse pancreas tissue stained with Collagen I antibody. IHC-P image submitted by a verified customer review.

Human lung tissue section stained with Collagen I antibody at 1:400.

FFPE right lobe of the liver tissue section. A:Central Vein (CV) fibrosis, B: Non-fibrotic CV, C: Perisinusodial fibrosis, D: Non-fibrotic area, E: Protat tract fibrosis, F: Septal fibrosis (arrow). Antigen retrieval: not required. Primary antibody: Anti-collagen type I at 1:1250 for 4C for 24hr. Secondary antibody: Peroxidase biotin-streptavidin rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: Anti-collagen type I is intra and extracellular. Staining: 3.3'-diaminobenzidine tetrahydrochloride was used as the chromogen. Nuclei were counterstained purple with hematoxylin.

Analysis of FFPE human lung. Primary antibody: Collagen I 1:400. Secondary antibody: Peroxidase goat anti-rabbit at 1:10,000 for 45 min at RT. Localization: Strong staining was observed in the extracellular matrix of the lung. Epithelial cells were negative. Staining: antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.

Analysis of HRP conjugate of NB600-408. FFPE Human Skin at pH 6. Primary antibody: Collagen Type I antibody at 10 ug/mL for 1 h at RT. Localization: Collagen Type I is secreted in the extracellular matrix.

Analysis of HRP conjugate of NB600-408. FFPE Human Skin tissue at pH9. Primary antibody: Collagen Type I antibody at 10 ug/mL for 1 h at RT. Localization: Collagen Type I is secreted in the extracellular matrix. Staining: Collagen Type I as precipitated brown signal (A) with hematoxylin purple nuclear counterstain. With corresponding negative conrol (B).

Imaging of rat calvarial defect bone. IHC-P image submitted by a verified customer review.

Immunohistological staining of EG treated rat kidneys for Collagen. Magnification X40, Scale bar 100 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0185009), licensed under a CC-BY license.

Analysis of Collagen I Antibody on alcohol-fixed rat lung tissue. Primary antibody dilution 1:200. Image from verified customer review.

Immunohistochemistry of Collagen I antibody. Tissue: human lung. Fixation: formalin fixed paraffin embedded. Antigen retrieval: user optimized. Primary antibody: Collagen 1 1:400 Secondary antibody: Peroxidase goat anti-rabbit at 1:10,000 for 45 min at RT. Localization: Strong staining was observed in the extracellular matrix of the lung. Epithelial cells were negative. Staining: antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.

Immunohistochemistry of Rabbit Anti-collagen type I antibody. Tissue: right lobe of the liver section. A:Central Vein (CV) fibrosis, B: Non-fibrotic CV, C: Perisinusodial fibrosis, D: Non-fibrotic area, E: Protat tract fibrosis, F: Septal fibrosis (arrow). Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Anti-collagen type I at 1:1250 for 4C for 24hr. Secondary antibody: Peroxidase biotin-streptavidin rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: Anti-collagen type I is intra and extracellular. Staining: 3.3'-diaminobenzidine tetrahydrochloride was used as the chromogen. Nuclei were counterstained purple with hematoxylin.

Immunohistochemistry-Paraffin: Rabbit Polyclonal Collagen I Antibody [NB600-408] - Collagen I was stained in human FFPE tissue. Primary antibody dilution: 1/1000 in blocking buffer. HIER antigen retrieval at pH 9 for 20min. AF750 conjugated version of the antibody was used (Catalog # NB600-408AF750). Image from a verified customer review.

Effects of fasudil treatment on tumor stroma in KPC mice.A) Pancreatic tumor tissues from vehicle, gemcitabine, or combination of gemcitabine plus fasudil treated KPC mice were harvested and stained for various stromal markers. Representative images are shown of H&E staining, alpha -SMA, Desmin, CD31, Collagen I, and Movat's pentachrome staining.

Western blot analysis. Comparison of collagen 1 (A and B) and collagen 2 (D and E) among the anterior horn, the body and the posterior horn: (A and D) young model; (B and E) adult model. Comparison of collagen 1 (C) and collagen 2 (F) content between the young and the adult specimens. Values with different superscripts differ for P < 0.01 (A,B). (G) Representative Western blot image for collagen 1, collagen 2 and GAPDH. N/group = 8.

EPS15 is targeted via a SPOP/SPOPL binding consensus motif.(A) Cartoon of human EPS15 domain-organization and the amino-acid sequence. Indicated by color code are the SPOP/SPOPL binding site (red) and the lysine residue (yellow), which is ubiquitinated in a CRL3SPOPL–dependent manner in vivo. In addition, the amino-terminal Ca2+-binding EF-hand motifs (EH), the coiled-coil domain involved in dimerization and the two carboxy-terminal ubiquitin-interacting motifs (UIMs) involved in ubiquitin-binding are indicated. (B) EPS15 ubiquitin-profiling. Peptides containing EPS15 modification sites were quantified with LC-MS/MS after enrichment of the K-epsilon -GG motif from whole cell digests of HeLa cells treated with siSPOPL or siControl. Normalized precursor mass intensity profiles for EPS15 sites corresponding to K793, K801 and K693 are shown (raw data in Figure 4—figure supplement 1B). Quantification of the beta -Actin K113 and the polyubiquitin K11 linkage peptide control for comparable enrichment. Data are mean ± SD, N = 3. **p≤0.01. (C) Purified SPOPL was incubated as indicated with GST-tagged wild-type EPS15 or GST-EPS15 mutants, where the predicted SPOPL binding motifs have been mutated individually (GST-EPS15S605-S607A and EPS15S744-S746A, respectively), pulled down with glutathione sepharose (IP [GST]) and bound proteins were analyzed by Coomassie blue staining (upper panel) and immunoblotting (lower panels). Note that SPOPL readily binds to GST-EPS15 and GST-EPS15S605-S607A, but this interaction is strongly reduced with the GST-EPS15S744-S746A mutant. (D) HeLa cells stably expressing GFP-tagged wild-type EPS15, the EPS15S744-S746A or the EPS15K793R mutants from a doxycycline-inducible promoter were transfected as indicated (+) with control siRNA or siRNA depleting SPOPL. The levels of EPS15-GFP, EGFR and for control tubulin (TUB) were analyzed by immunoblotting with specific antibodies. Experiments were quantified in Fiji and the EPS15 levels plotted as fold-increase compared to controls. Data are mean ± SEM, N = 4. *p≤0.05. Note that SPOPL depletion does not further increase the levels of both EPS15 mutants. (E) Total cell extracts were prepared from HeLa cells expressing either GFP-tagged wild-type, the EPS15S744-S746A mutant or the EPS15K793R mutant in the presence (+) or absence (-) of HA-tagged SPOPL overexpression. The levels of EPS15-GFP, SPOPL-HA and control GAPDH were analyzed by immunoblotting. Note that overexpression of SPOPL-HA is able to induce degradation of wild-type but not the EPS15S744-S746A-GFP or the EPS15K793R-GFP mutant.DOI://dx.doi.org/10.7554/eLife.13841.009EPS15 is targeted via a SPOP/SPOPL binding consensus motif.(A) Alignments of the carboxy-terminal domains of EPS15 proteins from various species. Conserved SPOPL-binding motifs and putative ubiquitination sites are highlighted by yellow boxes. (B) Peptides containing EPS15 modification sites were quantified with LC-MS/MS after enrichment of the K-epsilon -GG motif from whole cell HeLa digests treated with siSPOPL and siControl. Raw intensities for each of the triplicate LC-MS/MS runs are shown with each of the siControl conditions scaled to 100% intensity. Normalized precursor mass intensity profiles for EPS15 sites corresponding to K793, K801 and K693 are shown, with only K793 showing significant downregulation in the depletion condition. Quantification of a peptide corresponding to beta -Actin K113 and the poly-ubiquitin K11 linkage peptide is also shown to demonstrate that enrichment variations did not influence the quantification of the EPS15 sites. Additionally, the total ion chromatographic intensities for each run are plotted to provide insight into the consistency of each of the separate experiments performed on different days. Data are mean ± SD, N = 3. (C) HeLa cell lines stably expressing wild-type EPS15-GFP, the EPS15S744-746A-GFP mutant or the EPS15K793R-GFP mutant from the inducible doxycycline-promoter were treated with doxycycline for 3 days, and analyzed by live cell imaging. Displayed are maximal projections of Z-stack acquisitions, fully covering cell height. Scale bar = 10 μm.DOI://dx.doi.org/10.7554/eLife.13841.010 Image collected and cropped by CiteAb from the following open publication (//elifesciences.org/articles/13841), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Local release of anti-IL-6 from IMB suppresses LN fibrosis. The skin allografts were harvested at 7 days post-transplantation. (A-i) Lymphatic vessel expansion (Lyve-1+) & HEV elongation (MECA79+) were similar between all groups. (A-ii,iii) Dense staining of collagen I & PDPN was seen in DLNs harvested from control & GelMA group compared to those from GelMA/anti-IL-6 group. (representative images from 4 different mice per group). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31024011), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Phf2 protects the liver from high fat & high sucrose diet-induced fibrogenesis during obesity. GFP or Phf2 were overexpressed through AAV strategy specifically in the liver of 7-week-old male C57Bl/6 J mice. Mice were then fed with either a chow diet (NCD) or with a high fat & high sucrose diet (HFHSD) for 16 weeks. Mice were studied in the fed state. a Representative western blot analysis showing the contribution of Phf2 overexpression to the regulation of the PI3K/Akt signaling & pro-fibrogenic pathways in liver extracts (n = 15 per group). b (Left) Oral glucose tolerance test & insulin levels during the OGTT test. (right) Insulin tolerance test (n = 15 per group). c Liver sections stained with hematoxylin & eosin (H&E), trichrome masson & sirius red are shown. Scale bars = 100 μm (n = 10 per group). d Percentage of fibrotic area & percentage of apoptotic hepatocytes (n = 15 per group). e Relative oxidized GSSG content & measurement of Gpx activity (n = 10 per group). f Levels of carbonylated proteins (n = 10 mice per group). All error bars represent mean ± SEM. Statistical analyses were made using unpaired t-test (b) or Anova, followed by Bonferonni’s test (d, e). *P < 0.01 HFHSD/GFP compared to HFHSD/Phf2, $P < 0.01 HFHSD/GFP compared to NCD/GFP, £P < 0.01 HFHSD/Phf2 compared to HFHSD/GFP Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-04361-y), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Collagen I Antibody [NB600-408] - alpha -SMA, LOX, & COL1A1 immunostaining in human thyroid cancers. Representative thyroid tumors serial sections from two different patients stained by IHC for alpha -SMA, COL1A1, & LOX protein expression & localization. In the top panel, the tumor edge/invasive front is specifically shown (scale bar 500 µm), while lower panel has higher magnification (scale bar 200 µm). * clusters of tumor invading cells detaching from the principal tumor mass (T). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31906302), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Collagen I Antibody [NB600-408] - Increased co-localization of collagen type I with ER stress marker, KDEL in post-mortem human glaucomatous TM tissues. Post-mortem human TM tissues from age-matched normal (n = 3) & glaucoma (n = 3) were stained with Col I (green) & KDEL (red) & co-localization was examined in anterior segment tissues. Representative images are shown at lower (A) scale bar is 100 microns) & higher (B) scale bar is 25 microns) magnification. Both Col I & KDEL was increased in human glaucomatous TM tissues. In addition, increased co-localization of Col I with KDEL was observed in human glaucomatous TM tissues. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-14938-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Collagen I Antibody [NB600-408] - Expression of osteoblastic differentiation markers in Human Gingival Mesenchymal Stem Cells. Adherent HGMSCs were cultured for 21 days in control medium or in differentiation medium (Diff) supplemented or not with 1 μM resveratrol (RV). At the end, cell homogenates were processed for western blotting analysis of the expression of the osteoblastic transcription factor RUNX-2 & of the osteogenic differentiation markers COL1A1, OPN & OCN. Densitometry of the specific bands (average ± S.D.) of three independent experiments is shown in the histograms Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31426798), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Collagen I Antibody [NB600-408] - Dex induced ECM synthesis is associated with ER stress in TM cells. Primary human TM cells (n = 3 strains) were treated with Veh or Dex (100 nM) for 3 days & stained for KDEL & fibronectin (A) or Col I (B). A prominent co-localization of fibronectin or Col I with KDEL was observed in Dex-treated TM cells indicating induction of ER stress & excessive intracellular ECM overload in primary TM cells. (C) GTM3 cells were treated with Veh or Dex for 3 days & total lysates were subjected to immunoprecipitation using KDEL antibody & immunoblotted for ECM proteins, GRP78 & GRP94. IgG antibody was used as a negative control. Total cellular lysates were used as input. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-14938-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Collagen I Antibody [NB600-408] - Dex increases Col I in the TM: Dex increases Col I & alpha -smooth muscle actin in the TM of Dex-treated mice (n = 3) compared with Veh-treated mice (n = 3). White box indicates TM. CB = ciliary body, I = Iris, C = cornea. Scale bar = 50 μm. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-14938-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Collagen I Antibody [NB600-408] - Spautin-1 prevents the expression of osteoblastic differentiation markers in Human Gingival Mesenchymal Stem cells. HGMSCs were cultured for 21 days in control medium or in differentiation medium supplemented with 1 μM resveratrol (Diff + RV) in the absence or in the presence of spautin-1 (Sp1). At the end, cell homogenates were processed for western blotting analysis of the expression of the osteoblastic transcription factor RUNX-2 & of the osteogenic differentiation markers COL1A1, OPN & OCN. Densitometry of the specific bands (average ± S.D.) of three independent experiments is shown in the histograms Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31426798), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Collagen I Antibody [NB600-408] - Effects of fasudil treatment on tumor stroma in KPC mice.A) Pancreatic tumor tissues from vehicle, gemcitabine, or combination of gemcitabine plus fasudil treated KPC mice were harvested & stained for various stromal markers. Representative images are shown of H&E staining, alpha -SMA, Desmin, CD31, Collagen I, & Movat's pentachrome staining. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28841710), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Collagen I Antibody [NB600-408] - Immunofluorescence staining of osteoblastic differentiation markers in Human Gingival Mesenchymal Stem Cells. HGMSCs were plated on sterile coverslips, let adhere & cultured for 21 days in control medium or in differentiation medium (Diff) supplemented or not with 1 μM resveratrol (RV). At the end, the coverslips were fixed & processed for immunofluorescence staining of the osteogenic differentiation markers. Fluorescence staining was quantified with the ImageJ software Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31426798), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Collagen I Antibody [NB600-408] - Collagen secretion defects in fibroblasts from patient 1a. (a) Control or patient fibroblasts incubated for 30 min to promote procollagen secretion were immunolabeled to detect COL1A1 (red) & in green, either Sec31A (to mark COPII‐coated ER exit sites) or GM130 (to mark the Golgi apparatus). (b) Extracellular collagen matrix was labeled to detect COL1A1. (c) Quantification of the mean Corrected Total Cell Fluorescence (CTCF) for COL1A1. Points show mean intensity from three independent experiments. Bars show standard deviation. (d) Immunoblot for COL1A1 in media (M) or lysates (L) of control or patient fibroblasts incubated in the presence of absence of ascorbate. (e) Unbiased proteomics was used to measure extracellular matrix collagens secreted from control or patient fibroblasts. Results from three independent experiments are shown. A ratio <1 indicates reduced collagen secretion from patient cells Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31568717), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Collagen I Antibody [NB600-408] - The effects of PA on TGF-beta 1-reduced PPAR gamma expression in HSC-T6 cells. (A,B) The effect of PA-induced PPAR gamma expression in rat hepatic stellate cells (HSCs). Cells were treated with PA (2 μM) for 0, 6, 12, & 24 h or PA (0.5, 1, & 2 μM) for 24 h, & PPAR gamma expression was analyzed by Western blotting. The effect of PA on TGF-beta 1-reduced PPAR gamma expression in rat HSCs; (C) cells were pretreated with various concentrations of PA (0.5, 1, & 2 μM) for 1 h, & then stimulated with TGF-beta 1 (5 ng/mL) for 24 h. Total protein extracted from cells was subjected to Western blotting to determine PPAR gamma expression; (D,E) cells were treated with TGF-beta 1 (5 ng/mL) for 24 h in the presence of PA and/or GW9662 (10 μM). Total protein extracted from cells was subjected to Western blotting to determine the expression of PPAR gamma & alpha -SMA & collagen I alpha 1. Protein bands were imaged using densitometry & analyzed using ImageJ software. The relative expression levels of target proteins were normalized using beta -actin as an internal control. The results are presented as the means ± SD of three independent experiments. # Significantly different from the control (p < 0.01). * Significantly different from the TGF-beta 1-treated group (p < 0.01). + Significantly different from the PA-treated group (p < 0.01). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31795488), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Collagen I Antibody [NB600-408] - The effects of PA on TGF-beta 1-induced alpha -SMA & collagen I alpha 1 expression via suppression of the Akt & MAPK signal pathways in HSC-T6 cells. (A,B) The inhibitory effects of PA on TGF-beta 1-induced alpha -SMA & collagen I alpha 1 expression via suppression of Akt & MAPK in rat hepatic stellate cells (HSCs); (A) cells were treated with TGF-beta 1 (5 ng/mL) for 24 h in the presence of LY294002 (LY; 10 μM), PD98059 (PD; 20 μM), SB203580 (SB; 20 μM), or SP600125 (SP; 20 μM); (B) cells were treated with TGF-beta 1 (5 ng/mL) for 24 h in the presence of PA & LY294002 (LY; 10 μM), PD98059 (PD; 20 μM), SB203580 (SB; 20 μM), or SP600125 (SP; 20 μM). Total protein extracted from cells was subjected to Western blotting to determine alpha -SMA & collagen I alpha 1 expression. Protein bands were imaged using densitometry & analyzed using ImageJ software. The relative expression levels of target proteins were normalized using beta -actin as an internal control. The results are presented as the means ± SD of three independent experiments. # Significantly different from the control (p < 0.01). * Significantly different from the TGF-beta 1-treated group (p < 0.01). + Significantly different from the PA-treated group (p < 0.01). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31795488), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Collagen I Antibody [NB600-408] - Cells expressing IL-1⍺ & IL-1R are observed in the fibrotic liver environment. (a) Cytokines in tissue lysates from mice at 9 wpi were measured by ELISA in liver. Data are presented as fold change relative to the mean of uninfected levels. N = 11–12 mice per group, pooled from three independent experiments. (b) IL-1 alpha levels in the sera at 9 wpi measured by ELISA. N = 9–14 mice per group, pooled from four independent experiments. (c) Immunofluorescence labeling of nuclei (DAPI white), IL-1⍺ (green), CD45 (red), & collagen1⍺1 (blue) in the liver of UI or 9 wpi WT mice. Number of cells staining positive for CD45 and/or IL-1⍺, average 2–3 fields of view where immune infiltrate was present from 3 mice per condition are quantified on the right. Error bars are standard deviation. (d) Immunofluorescent labeling of nuclei (DAPI white), ⍺-smooth muscle actin (green), IL-1R (red), & collagen1⍺ (blue) in the liver of uninfected or 9 wpi WT. Inset, arrow head represents ⍺-smooth muscle actin/IL-1R co-staining cells (arrow heads). Number of cells staining positive for IL-1R and/or ⍺-SMA, average of 4–8 fields of view from 3 mice are quantified on the right. Error bars are standard deviation. (c,d) represent maximum intensity projections of 9–13 μm thick z-stacks. Scale bar represents 50 μm. Error bars are standard error of the mean except where noted otherwise. *P < 0.05; **P < 0.01; ***P < 0.001 by unpaired Student’s t test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32973293), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: Collagen I Antibody [NB600-408] - Characterization of decellularized heart matrix for DNA & extracellular matrix content with DNA quantification, histological staining, & multiphoton imaging. (A) DNA content of the right ventricle (RV, triangles), septum (ST, circles), & left ventricle (LV, squares) for cadaveric hearts compared to decellularized Langendorff (n = 4) or 4-Flow cannulated hearts (n = 4). (B) Masson’s trichrome staining of representative Langendorff (n = 4) & 4-Flow (n = 4) hearts for preserved collagen (blue) & clearance of nuclear remnants (absence of positively stained purple nuclei). (C) Multiphoton microscopy visualization of collagen (pseudocolored - cyan) & elastin (pseudocolored - yellow) fibrillar strcutures & organization in 3D (n = 4). (D) Immunostaining of histology sections important extracellular matrix proteins: collagen I, collagen IV, & laminin (n = 4). (E) The list of preserved extracellular matrix proteins in decellularized 4-Flow hearts as detected by mass spectrometry (n = 4). Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-25883-x), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Collagen I Antibody [NB600-408] - PARP1 regulates VSMCs osteogenic transition, not Na/P co-transporter. a, b Quantitative RT-PCR (a) & western blot (b) analysis of PIT1 & PIT2 expression in VSMCs, which were infected with Scrambled (Scr shRNA) or PARP1 shRNA, & treated with high Pi. c–e Rat VSMCs were infected with adenovirus carrying Scr shRNA, PARP1 shRNA, Null or PARP1 gene for 48 h & then incubated with osteogenic media for 14 days. PARP1 inhibitors 3AB (10 mM) & PJ34 (10 μM) were added every day. Western blot analysis & related quantification of osteogenic-related marker genes (OPN, ColIA1, & OC) & smooth muscle marker genes (SMA, SM22 & SMMHC) expression in VSMCs with PARP1 knockdown (c), PARP1 inhibitors 3AB & PJ34 (d), & PARP1 overexpression (e). (n = 5 for each group). Statistical significance was assessed using one-way ANOVA for multiple comparison & two-tailed t-tests for two groups & is presented as follows: NS: no significance, **P < 0.01 & ##P < 0.01. All values are means ± S.D. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30867423), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Collagen I Antibody [NB600-408] - CM derived from OS cells stimulates the BM‐MSCs trans‐differentiation into CAF‐like cells. (A) FACS analysis of NG‐2 & CD31 expression in BM‐MSCs treated for 48 h with CM OS cells. (B) BM‐MSCs were stimulated for 48 h with CM from OS cells & alpha ‐SMA & Col I‐ alpha 1 expression was assessed by immunoblot analysis. Results are representative of four biological replicates. (C) Collagen contraction assay of BM‐MSCs treated for 24 h with St Med or HOS CM. Data are expressed as percentages of the relative area of collagen disc following contraction in comparison with an empty well (mean ± SD, n = 3 biological replicates performed in duplicate). * P < 0.05 vs. St Med. (D) Migration assay of HOS cells stimulated for 48 h with CM derived from BM‐MSCs previously activated or not activated by tumour cells (CM BM‐MSCs St/HOS). Cells were allowed to migrate toward complete medium (FBS 10%). Untreated cells (St Med) were used as control. Results are the mean ± SEM of three biological replicates. *** P < 0.001 vs. St Med. (E) Invasion & transmigration (F) assays of HOS cells treated as in (D) (mean ± SD, n = 3 independent experiments). ** P < 0.005 vs. St Med; *** P < 0.001 vs. St Med. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29517849), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Collagen I Antibody [NB600-408] - Human PRP infusion improves regeneration of the damaged endometrium & down-regulates expression of fibrosis-related factors. (A) Design of Experiment 1. In AS mice, PRP was injected only into the right horn at 7 days after inducing injury to the bilateral uterine horns. At 14 days, all mice were sacrificed & the uterine horns were prepared for analysis. (B) Hematoxylin & eosin staining of the endometrial tissues to evaluate morphologic structures in the PRP-treated horn of AS mice. (C) Masson’s trichrome staining to evaluate collagen deposition (blue) in the PRP-treated horns of AS mice. (D) Immunofluorescence staining of collagen type 1A (COL1A1) in the PRP-treated horn of AS mice. Green & red indicate COL1A1 & nuclei, respectively. (E) RT-qPCR & (F) real-time RT-qPCR analyses of expression of fibrosis-related factors in the PRP-treated horn of AS mice. Sham, n = 4; AS or PRP-treated AS, n = 5. *p < 0.01; Scale bar: 100 & 200 μm. AS, Asherman’s syndrome; PCR, polymerase chain reaction; PRP, platelet-rich plasma; RT-qPCR, reverse transcription-quantitative PCR. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32116803), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Collagen I Antibody [NB600-408] - Fibroblast-derived CDM induces sustained growth inhibition of cancer cells.(a) Collagen I & fibronectin staining of CDM generated by NFs (TIFFs). Scale bar, 20 μm. (b) Proliferation of MDA-MB-231 & HeLa cells plated on TIFF CDM or on plastic in full medium for the indicated times. n(HeLa)=10, n(MDA-MB-231)=8. (c) Proliferation of MDA-MB-231 & HeLa cells after detachment from TIFF matrices (6 days on matrix before detachment) & replating on plastic in full medium for the indicated times n(HeLa)=7, n(MDA-MB-231)=10. (d) Representative images of MDA-MB-231 cell morphology on CDM & plastic. Shown are maximum intensity projections of confocal images. Scale bar, 10 μm. (e) Schematic representation of the experimental set-up. Red arrows indicate the time points of sample collection for Illumina gene expression analysis. (f) Common gene expression changes in MDA-MB-231 & HeLa cells on CDM & 5 days after CDM detachment (CDM to plastic (pl.)). The numbers of commonly regulated genes (up- or downregulated) in both cell lines are indicated in the table. Upregulated genes are marked with red & downregulated genes with blue. All data are mean±s.e.m. Unpaired t-test was used for statistical analyses. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27488962), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Collagen I Antibody [NB600-408] - Spautin-1 prevents the autophagy-associated osteoblastic differentiation of Human Gingival Mesenchymal Stem cells. a HGMSCs were plated on sterile coverslips, let adhere & cultured for 21 days in control medium or in differentiation medium supplemented with 1 μM resveratrol (Diff + RV) in the absence or the presence of spautin-1 (Sp1). The coverslips were then fixed & processed for immunofluorescence staining of the autophagy marker LC3 & of the osteoblastic differentiation marker OCN. As an index of autophagy in the cells, LC3 puncta were quantified (as per the guidelines 36). b HGMSCs were plated & treated as in a & the coverslips processed for immunofluorescence staining of the osteoblastic differentiation markers OCN & COL1A1. Integrated fluorescence intensity quantified with the ImageJ software is shown in the histograms. c HGMSCs cells were plated on plastic & treated as in a & at the end the cultures were processed for Alizarin Red S staining of extracellular calcium deposits. The stained area was quantified using the ImageJ software Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31426798), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Collagen I Antibody [NB600-408] - Collagen secretion defects in fibroblasts from patient 1a. (a) Control or patient fibroblasts incubated for 30 min to promote procollagen secretion were immunolabeled to detect COL1A1 (red) & in green, either Sec31A (to mark COPII‐coated ER exit sites) or GM130 (to mark the Golgi apparatus). (b) Extracellular collagen matrix was labeled to detect COL1A1. (c) Quantification of the mean Corrected Total Cell Fluorescence (CTCF) for COL1A1. Points show mean intensity from three independent experiments. Bars show standard deviation. (d) Immunoblot for COL1A1 in media (M) or lysates (L) of control or patient fibroblasts incubated in the presence of absence of ascorbate. (e) Unbiased proteomics was used to measure extracellular matrix collagens secreted from control or patient fibroblasts. Results from three independent experiments are shown. A ratio <1 indicates reduced collagen secretion from patient cells Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31568717), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Collagen I Antibody [NB600-408] - The effects of PA on TGF-beta 1-induced alpha - SMA & collagen I alpha 1 expression in HSC-T6 cells. (A,B) The inhibitory effect of PA on TGF-beta 1-induced alpha -SMA & collagen type I mRNA & protein expression in rat hepatic stellate cells. Cells were pretreated with 0.5, 1, & 2 μM PA for 1 h, & then stimulated with TGF-beta 1 (5 ng/mL) for 24 h. Total RNA extracted from cells was analyzed by the real-time polymerase chain reaction to determine alpha -SMA & ColIa1 mRNA expression; (C) the total protein extracted from cells was subjected to Western blotting to determine alpha -SMA & collagen I alpha 1 expression. Protein bands were imaged using densitometry & analyzed using ImageJ software. The relative expression levels of target proteins were normalized using beta -actin as an internal control. The results are expressed as the means ± SD of three independent experiments. # Significantly different from the control (p < 0.01). * Significantly different from the TGF-beta 1-treated group (p < 0.01). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31795488), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Frozen: Collagen I Antibody [NB600-408] - Cryopreserved murine brain tissue stained with Collagen I Antibody [Alexa Fluor® 647] (Catalog # NB600-408AF647) (red) and Hoechst (blue). Image from a verified customer review.

Reactivity:

Hu, Mu, Rt, Po, Bv, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, 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	NB600-408AF350
Alexa Fluor 405	NB600-408AF405
Alexa Fluor 488	NB600-408AF488
Alexa Fluor 532	NB600-408AF532
Alexa Fluor 594	NB600-408AF594
Alexa Fluor 647	NB600-408AF647
Alexa Fluor 700	NB600-408AF700
Alexa Fluor 750	NB600-408AF750
Allophycocyanin	NB600-408APC
Biotin	NB600-408B
CoraFluor 1	NB600-408CL1
DyLight 350	NB600-408UV
DyLight 405	NB600-408V
DyLight 488	NB600-408G
DyLight 550	NB600-408R
DyLight 594	NB600-408DL594
DyLight 650	NB600-408C
DyLight 680	NB600-408FR
DyLight 755	NB600-408IR
FITC	NB600-408F
HRP	NB600-408H
Janelia Fluor 525	NB600-408JF525
Janelia Fluor 549	NB600-408JF549
Janelia Fluor 585	NB600-408JF585
Janelia Fluor 635	NB600-408JF635
Janelia Fluor 646	NB600-408JF646
Janelia Fluor 669	NB600-408JF669
PE	NB600-408PE
PerCP	NB600-408PCP

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DY453

Mouse CXCL1/KC DuoSet ELISA, 15 Plate

(290 Publications)

1 Kit (for 5 Plates)

DY453

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

Applications:

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NB100-690

alpha Tubulin Antibody (DM1A) - BSA Free

(14 Reviews)

(146 Publications)

0.1 ml

NB100-690

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

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

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

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

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

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

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

Reactivity:

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

Species Glossary

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

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-690AF350
Alexa Fluor 405	NB100-690AF405
Alexa Fluor 488	NB100-690AF488
Alexa Fluor 532	NB100-690AF532
Alexa Fluor 594	NB100-690AF594
Alexa Fluor 647	NB100-690AF647
Alexa Fluor 700	NB100-690AF700
Alexa Fluor 750	NB100-690AF750
Allophycocyanin	NB100-690APC
Biotin	NB100-690B
CoraFluor 1	NB100-690CL1
DyLight 350	NB100-690UV
DyLight 405	NB100-690V
DyLight 488	NB100-690G
DyLight 550	NB100-690R
DyLight 594	NB100-690DL594
DyLight 650	NB100-690C
DyLight 680	NB100-690FR
DyLight 755	NB100-690IR
FITC	NB100-690F
HRP	NB100-690H
Janelia Fluor 525	NB100-690JF525
Janelia Fluor 549	NB100-690JF549
Janelia Fluor 585	NB100-690JF585
Janelia Fluor 635	NB100-690JF635
Janelia Fluor 646	NB100-690JF646
Janelia Fluor 669	NB100-690JF669
PE	NB100-690PE
PerCP	NB100-690PCP
mFluor Violet 450 SE	NB100-690MFV450
mFluor Violet 500 SE	NB100-690MFV500
mFluor Violet 610 SE	NB100-690MFV610

Available Formulations

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

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MKC00B

Mouse CXCL1/KC Quantikine ELISA Kit, 2 Plate

(199 Publications)

1 Kit

MKC00B

STAT3 regulates chemoattractive CXCL1 expression.(a) ELISA of CXCL1 levels in lung lysates and BAL at 6 and 13 weeks post AdCre. Data were analysed by Student’s t-test and shown as mean±s.d., n≥7 mice per genotype and time point. (b) Representative in situ hybridization with probes specific for murine Cxcl1 (AS) in lung tumours at 13 weeks post AdCre. Sense probes (S) were used as negative control and surfactant protein C (SP-C) as lung epithelial maker. Scale bar, 100 μm. (c) Kaplan–Meier plot showing overall survival of lung adenocarcinoma patient samples harbouring smoking history with high or low IL-8 mRNA expression (expression values of upper quartile, log-rank test) n≥35–104 per group. (d) Human A549 lung cancer cell line (KRASG12S mutated) was stimulated with designated cytokines for indicated time points. IL-8 or CXCL1 expression was analysed via qRT–PCR. Data were analysed by one-way ANOVA with Tukey’s multiple comparison test and shown as mean±s.d. (e) A549 cells transducted with lentiviral vectors expressing nonspecific scrambled small hairpin (sh)RNA or shRNA against STAT3 were stimulated with indicated cytokines for 60 min. Data were analysed by one-way ANOVA with Tukey’s multiple comparison test and shown as mean±s.d. (f) Primary pneumocytes were isolated and infected with AdCre. 120 h post infection cells were stimulated with indicated cytokines for 60 min. Cxcl1 expression was analysed by qRT–PCR. Data are shown as mean±s.e.m. (g) Murine 3T3 fibroblasts and STAT3 overexpressing 3T3 fibroblasts were stimulated as in f. Cxcl1 expression was analysed by qRT–PCR. Data were analysed by one-way ANOVA with Tukey’s multiple comparison test and shown as mean± s.d. Values in d–g are presented as fold change of relative mRNA expression compared with each unstimulated individual cell line. At least two independent experiments with three individual plates per stimulation were performed. For all graphs: *P<0.05; **P<0.01; ***P<0.001. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms7285), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

Applications:

Conjugate	Catalog #	Availability	Size	Price
HRP	SMKC00B
HRP	PMKC00B

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AF2645

GFR alpha-3/GDNF R alpha-3 Antibody [Unconjugated]

(16 Publications)

100 μg

AF2645

GFRa‑3/GDNF Ra‑3 was detected in immersion fixed frozen sections of mouse embryo (15 d.p.c.) using 15 µg/mL Goat Anti-Mouse GFRa‑3/GDNF Ra‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2645) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to developing dorsal root ganglion and spinal cord. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

,

Applications: ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF2645

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF2645-SP

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NB120-13888

DNMT3A Antibody (64B1446) - BSA Free

(3 Reviews)

(124 Publications)

0.1 mg

NB120-13888

Immunohistochemistry-Paraffin: DNMT3A Antibody (64B1446) [NB120-13888] - Detection of DNMT3A on human hepatocellular carcinoma tissue section (NBP2-30221) using 1:100 dilution of DNMT3A antibody (clone 64B1446). The antibody generated a specific cytoplasmic staining in all the cancer cells while some of the cells depicted nuclear staining also.

Immunocytochemistry/Immunofluorescence: DNMT3A Antibody (64B1446) [NB120-13888] - Expression of Dnmt3a in the nuclei of post-mitotic neurons in the olfactory epithelium (OE). DNMT3a (red) is co-expressed with neuron-specific tubulin (green) throughout the development of the olfactory epithelium. Data courtesy of A. Jane Roskams, University of British Columbia.

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - Intracellular flow cytometric staining of 1 x 10^6 CHO (A) and HEK-293 (B) cells using Dnmt3a antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1x10^6 cells was used.

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - Using the PE direct conjugate, an intracellular stain was performed on NTERA-2 cells with DNMT3A (64B1446) NB120-13888PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.

Immunocytochemistry/Immunofluorescence: DNMT3A Antibody (64B1446) [NB120-13888] - DNMT1 reveals a SCMC-like cortical localization with aberrant localization in Nlrp2tm1a/tm1a derived oocytes. Whole mount immunofluorescence for DNMT3A in unfertilized control oocytes reveals a characteristic metaphase associated localization and no difference is noted in oocytes derived from Nlrp2tm1a/tm1adams. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep44667), licensed under a CC-BY license.

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on Jurkat cells with DNMT3A (64B1446) NB120-13888AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

<b>Biological Strategies Validation. </b>Western Blot: DNMT3A Antibody (64B1446) [NB120-13888] - NTERA-2 cells were treated with Zebularine as indicated for 24 hours. Cell lysates were prepared and separated on a 7.5% gel by SDS-PAGE. Protein was transferred to PVDF membrane and blocked in 5% non-fat milk. The membrane was probed with 2 ug/mL anti-Dnmt3a in 1% milk, and detected with an anti-mouse HRP secondary antibody using chemiluminescence. Note the decrease in Dnmt3a expression upon treatment with 500 uM Zebularine (arrowhead). Additional bands at 90 and 75 kDa can also be detected with this antibody and may represent alternative splice variants. Tubulin is shown as a loading control.

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on HepG2 cells with DNMT3A (64B1446) NB120-13888APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to allophycocyanin (APC).

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on HepG2 cells with DNMT3A (64B1446) NB120-13888AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on HeLa cells with DNMT3A (64B1446) NB120-13888APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to allophycocyanin (APC).

<b>Genetic Strategies Validation. </b>Knockdown Validated: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888] - DNMT3A knockdown increases miR‐200b expression via promoter demethylation. Western blot assay of DNMT3A expression in MDA‐MB‐231 and BT549 cells after transfection with DNMT3A siRNA (si‐DNMT3A) or negative control (NC). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30324719/) licensed under a CC-BY license.

Flow Cytometry: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888] - Protein levels of Dnmt3a & MeCP2 are reduced in inhibitory neurons of conditional knockout mice.(A) Western blot of half brain hemisphere from Dnmt3a or Mecp2 cKO mice demonstrating loss of each protein. (B) Immunofluorescence (IF) images of WT, Dnmt3a cKO or Mecp2 cKO mice probing for Dnmt3a or MeCP2 (red), the Sun1-sfGFP-myc-PA fusion protein that marks the nuclear envelope & is dependent on Cre expression (green), & DAPI to mark genomic DNA (blue). n = 3 mice per genotype (western), n = 3 mice (IF) representative images for two brain regions shown. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32159514), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-268A

Alt Catalog# IMG-268A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB120-13888AF350
Alexa Fluor 405	NB120-13888AF405
Alexa Fluor 488	NB120-13888AF488
Alexa Fluor 532	NB120-13888AF532
Alexa Fluor 594	NB120-13888AF594
Alexa Fluor 647	NB120-13888AF647
Alexa Fluor 700	NB120-13888AF700
Alexa Fluor 750	NB120-13888AF750
Allophycocyanin	NB120-13888APC
Biotin	NB120-13888B
CoraFluor 1	NB120-13888CL1
DyLight 350	NB120-13888UV
DyLight 405	NB120-13888V
DyLight 488	NB120-13888G
DyLight 550	NB120-13888R
DyLight 594	NB120-13888DL594
DyLight 650	NB120-13888C
DyLight 680	NB120-13888FR
DyLight 755	NB120-13888IR
FITC	NB120-13888F
HRP	NB120-13888H
Janelia Fluor 525	NB120-13888JF525
Janelia Fluor 549	NB120-13888JF549
Janelia Fluor 585	NB120-13888JF585
Janelia Fluor 635	NB120-13888JF635
Janelia Fluor 646	NB120-13888JF646
Janelia Fluor 669	NB120-13888JF669
PE	NB120-13888PE
mFluor Violet 450 SE	NB120-13888MFV450
mFluor Violet 500 SE	NB120-13888MFV500
mFluor Violet 610 SE	NB120-13888MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB120-13888SS
Azide and BSA Free	NBP2-80699

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NB600-506

alpha Tubulin Antibody (YL1/2) - BSA Free

(4 Reviews)

(57 Publications)

0.1 ml

NB600-506

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (YL1/2) [NB600-506] - NIH3T3 cells were fixed and permeabilized for 10 minutes with -20C MeOH. The cells were incubated with alpha Tubulin Antibody [YL1/2] (NB600-506) at 1ug/ml overnight at 4C and detected with an anti-rat DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (YL1/2) [NB600-506] - Rat FR cells were fixed and permeabilized for 10 minutes with -20C MeOH. The cells were incubated with alpha Tubulin Antibody [YL1/2] (NB600-506) at 1ug/ml overnight at 4C and detected with an anti-rat DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Western Blot: alpha Tubulin Antibody (YL1/2) [NB600-506] - Total protein from human HeLa, mouse 3T3, rat PC12 and African green monkey Cos7 cell lines was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with a 1:6000 dilution of anti-Tubulin (YL 1/2) in 1% non-fat milk in TBST and detected with an anti-rat HRP secondary antibody using chemiluminescence. Alpha tubulin molecular weight: 50 kDa.

Immunohistochemistry-Paraffin: alpha Tubulin Antibody (YL1/2) [NB600-506] - Tubulin antibody was tested in human breast cancer xenograft using DAB with hematoxylin counterstain.

Flow Cytometry: alpha Tubulin Antibody (YL1/2) [NB600-506] - An intracellular stain was performed on PC12 cells with Tubulin [YL1/2] Antibody NB600-506AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: alpha Tubulin Antibody (YL1/2) [NB600-506] - Tissue levels of VEGF and PCNA.VEGF levels were estimated by western blot. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0112995), licensed under a CC-BY license.

Western Blot: alpha Tubulin Antibody (YL1/2) [NB600-506] - Western blot analysis of alpha Tubulin (molecular weight: 50 kDa) expression in 1) HeLa, 2) NTERA-2, 3) A431, 4) HepG2, 5) MCF7, 6) NIH-3T3, 7) PC-12 and 8) Cos 7 whole cell lysates using NB600-506.

Western Blot: alpha Tubulin Antibody (YL1/2) [NB600-506] - Western Blot analysis of whole cell lysates of HeLa, NIH-3T3 and PC-12 cell lines using Tubulin antibody (clone YL1/2). Alpha tubulin molecular weight: 50 kDa.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (YL1/2) [NB600-506] - Tubulin YL1/2 antibody was tested in HeLa cells with Dylight 550 (red). Nuclei were counterstained with DAPI (blue).

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (YL1/2) [NB600-506] - B-Tubulin staining (488) and Pericentrin (594) in Mitotic Cell. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (YL1/2) [NB600-506] - IF Confocal analysis of NIH/3T3 cells using Tubulin antibody (NB600-506, 1:5). An Alexa Fluor 488-conjugated Goat to rat 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).

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (YL1/2) [NB600-506] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-Tubulin (YL1/2) at a 1:200 dilution overnight at 4C and detected with an anti-rat Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow Cytometry: alpha Tubulin Antibody (YL1/2) [NB600-506] - Analysis of Alexa Fluor (R) 647 conjugate of NB600-506. An intracellular stain was performed on HeLa cells with Tubulin antibody (YL1/2) NB600-506AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: alpha Tubulin Antibody (YL1/2) [NB600-506] - An intracellular stain was performed on NIH3T3 cells with Tubulin [YL1/2] Antibody NB600-506AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: alpha Tubulin Antibody (YL1/2) - BSA Free [NB600-506] - Tissue levels of VEGF & PCNA.VEGF levels were estimated by (A) western blot & (B) ELISA. (C) PCNA levels estimated by western blot. Data were normalized with tubulin. (D) Representative images of PCNA immunohistochemistry staining in the nucleus is shown for CS-exposed mice (CSE) & CS-exposed & LGF-treated mice (CSE+LGF). Scale bars = 50 µm. LGF promoted an increase of PCNA+ cells as showed in the bar graph. ** P<0.01 vs. air-exposed mice; ¥ P<0.05 vs. CS-exposed mice (n = 5 per group). Data are presented as mean ± SEM. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25401951), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: alpha Tubulin Antibody (YL1/2) - BSA Free [NB600-506] - LGF ameliorates oxidative stress.Bars represent (A) 3NT & (B) Nrf2 levels estimated by western blot. Data were normalized with tubulin. * P<0.05 vs. air-exposed mice; ¥ P<0.05 vs. CS-exposed mice (n = 5 per group). Data are presented as mean ± SEM. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25401951), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: alpha Tubulin Antibody (YL1/2) - BSA Free [NB600-506] - Role of cell-type-specific cyclooxygenase-2 (Cox-2) expression in Ras/p53-mediated tumor formation. a Experimental scheme. b Microscopic phenotype & histology of three-dimensional (3D) organoids with/without celecoxib treatment. c Relative 3D organoid formation was determined by the number of organoids larger than 100 µm, n = 3 independent experiments, 25 fields at high power field (HPF) per sample. Veh, vehicle control; C25, 25 µM; C50, 50 µM celecoxib. d Experimental scheme. e Ras/p53-mediated tumor incidence with/without condition Cox-2 knockout. Ptgs2wt/wt, n = 15 animals; Ptgs2flox/flox, n = 14 animals. f, g Histological phenotypes. h–k Immunoblotting of Keratin5 (Krt5), Krt6 & Loricrin (Lor) demonstrated decreased tumor susceptibility but increased differentiation status, n = 4 independent experiments. l Summary of the contribution of physiological stress factors in the susceptibility of tumor development from tumor-competent Krt15+ progenitors. Data with bar graphs are represented as mean ± SEM. Statistical significance was determined by pair-wise comparison using t test; ns = not significant, *p < 0.05, **p < 0.005. Scale bars, 100 µm. Ptgs2 prostaglandin-endoperoxide synthase 2 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31110179), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: alpha Tubulin Antibody (YL1/2) - BSA Free [NB600-506] - Microenvironmental acidic stressors & tumor susceptibility. a Experimental scheme. Epithelial cells isolated from the regions adjacent or distant from squamocolumnar junction (SCJ). b Microscopic phenotype of three-dimensional (3D) organoids from control & Krt15-CrePR; LSL-KrasG12D; p53flox/flox mice. Histology of 3D organoids demonstrated that oncogenic Ras/p53 increased abnormal growth features. c–e Quantification of relative organoid formation (n = 3 independent experiments, 25 fields at high power field (HPF) per sample) & size distribution (n ≥ 150) from control & experimental mice expressing oncogenic Ras/p53 combination. f Experimental scheme. Phosphate-buffered saline (PBS) (vehicle) or PPI were daily treated by intraperitoneal (i.p.) injections (5 consecutive days per week). g Tumor incidence from Krt15-CrePR; LSL-KrasG12D; p53flox/flox mice, with/without PPI treatment, n = 9 animals per each. Statistical significance was determined by Fisher’s exact test; *p < 0.05. h–j Immunoblotting of Keratin5 (Krt5) & Krt6 demonstrated relatively less tumor formation in the PPI treatment group, n = 5 independent experiments. k–m Immunostaining of Krt5, Krt6, & phospho-histone H3 (ph-H3), & histology demonstrated suppressed tumor formation by daily PPI treatment in Krt15-CrePR; LSL-KrasG12D; p53flox/flox mice. PPI, proton pump inhibitor. Data with bar graphs are represented as mean ± SEM. Statistical significance was determined by pair-wise comparison using t test; *p < 0.05, **p < 0.005. Scale bars, 100 µm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31110179), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Av, Ce, Dr, In, Ma, Pm, Pm, Ye

Species Glossary

Reactivity: Hu, Mu, Rt, Av, Ce, Dr, In, Ma, Pm, Pm, Ye

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Rat Monoclonal

Host: Rat Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-506AF350
Alexa Fluor 405	NB600-506AF405
Alexa Fluor 488	NB600-506AF488
Alexa Fluor 532	NB600-506AF532
Alexa Fluor 594	NB600-506AF594
Alexa Fluor 647	NB600-506AF647
Alexa Fluor 700	NB600-506AF700
Alexa Fluor 750	NB600-506AF750
Allophycocyanin	NB600-506APC
Biotin	NB600-506B
CoraFluor 1	NB600-506CL1
DyLight 350	NB600-506UV
DyLight 405	NB600-506V
DyLight 488	NB600-506G
DyLight 550	NB600-506R
DyLight 594	NB600-506DL594
DyLight 650	NB600-506C
DyLight 680	NB600-506FR
DyLight 755	NB600-506IR
FITC	NB600-506F
HRP	NB600-506H
Janelia Fluor 525	NB600-506JF525
Janelia Fluor 549	NB600-506JF549
Janelia Fluor 585	NB600-506JF585
Janelia Fluor 635	NB600-506JF635
Janelia Fluor 646	NB600-506JF646
Janelia Fluor 669	NB600-506JF669
PE	NB600-506PE
mFluor Violet 450 SE	NB600-506MFV450
mFluor Violet 500 SE	NB600-506MFV500
mFluor Violet 610 SE	NB600-506MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-506SS
Azide and BSA Free	NBP2-80571

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MMA00

Mouse CCL3/MIP-1 alpha Quantikine ELISA Kit, 2 Plate

(77 Publications)

1 Kit (2 Plates)

MMA00

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

Applications:

Conjugate	Catalog #	Availability	Size	Price
HRP	PMMA00
HRP	SMMA00

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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 Type

Preservative

Protein Or Peptide

Research Areas

Has Sample Size

Promotional Item

Species Glossary

IRE1 alpha [p Ser724] Antibody - BSA Free

Survivin Antibody - BSA Free

PGC1 alpha Antibody - BSA Free

LC3A Antibody - BSA Free

Collagen I Antibody

Mouse CXCL1/KC DuoSet ELISA, 15 Plate

alpha Tubulin Antibody (DM1A) - BSA Free

Mouse CXCL1/KC Quantikine ELISA Kit, 2 Plate

GFR alpha-3/GDNF R alpha-3 Antibody [Unconjugated]

DNMT3A Antibody (64B1446) - BSA Free

alpha Tubulin Antibody (YL1/2) - BSA Free

Mouse CCL3/MIP-1 alpha Quantikine ELISA Kit, 2 Plate