ELISA: Novus Biologicals

NB100-2323

IRE1 alpha [p Ser724] Antibody - BSA Free

(9 Reviews)

(356 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 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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DY485

Mouse IFN-gamma DuoSet ELISA, 15 Plate

(369 Publications)

1 Kit (for 5 Plates)

DY485

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

Applications:

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DLB50

Human IL-1 beta/IL-1F2 Quantikine ELISA Kit

(344 Publications)

1 Kit

DLB50

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Conjugate	Catalog #	Availability	Size	Price
HRP	PDLB50
HRP	SLB50

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NB600-408

Collagen I Antibody - BSA Free

(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 646	NB600-408JF646
Janelia Fluor 669	NB600-408JF669
PE	NB600-408PE
PerCP	NB600-408PCP

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-408-0.025mg

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NB100-449

HIF-1 alpha Antibody

(28 Reviews)

(316 Publications)

0.1 ml

NB100-449

Simple Western: HIF-1 alpha Antibody [NB100-449] - Simple Western lane view shows a specific band for HIF-1 alpha in 0.5 mg/ml of hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Mouse Brain, Neurons 40X

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1-alpha by immunohistochemistry. Sample: FFPE section of renal cell carcinoma. Antibody: Affinity purified rabbit anti-HIF1-alpha antibody (NB110-58773). Detection: DAB

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of Human HIF-1 alpha by Western Blot and Immunoprecipitation. Samples: Whole cell lysate (5, 15 and 50 ug for WB; 1 mg for IP, 20% of IP loaded) from HeLa cells that were either treated with cobalt chloride (+; 200 uM) or mock treated (-). Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody used for WB at 0.1 ug/mL(A) and 1 ug/mL (B) and used for IP at 3 ug/mg lysate. HIF-1 alpha was also immunoprecipitated by a previous lot of this antibody. Detection: Chemiluminescence with exposure times of 30 seconds (A) and 10 seconds (B).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Blot showing the effect of hypoxia on the protein expression levels of the purine biosynthetic enzymes. HIF-1 alpha is stabilized in hypoxia as expected, and no significant increase in the purine enzymes was detected between normoxic (21% oxygen) and hypoxic (1% oxygen) growth conditions. The positions of molecular markers surrounding each band of interest are shown for each blot. ADSL (NBP2-03107), ATIC (NBP2-01941), FGAMS (NBP1-84691), GART (H00002618-M01), HIF-1a (NB100-449), PAICS (NBP2-02817), PPAT (NBP2-02056). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32439803/) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Murine primary bone marrow derived macrophages stained with HIF1-alpha antibody (red). Nuclei were counterstained with DAPI (blue). Image from verified customer review.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB100-449] - Analysis of microvascular density and HIF-1alpha activity. Low, homogenous expression of HIF-1alpha was detected in the tumors of NNK treated mice (40X, scale bar = 25 um). Image collected and cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-11-4), licensed under a CC-BY license.

Flow Cytometry: HIF-1 alpha Antibody [NB100-449] - HeLa cells were treated for 15 hrs with 200 uM CoCl2, fixed in PFA, and permeabilized in 90% MeOH. 1 X 10^6 cells were stained with 0.125 ug anti-HIF-1 alpha and secondary FITC-conjugated goat anti-rabbit (in a 150 uL reaction). Black: treated, anti-KLH control IgG; Red: untreated, anti-HIF-1 alpha; Blue: treated, anti-HIF-1 alpha.

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of HIF-1 alpha in human myeloma cell lysate using anti-HIF-1 alpha. Cells were untreated or treated with IGF-1, IL-6 or CoCl2. Image from verified customer review.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of HIF-1 alpha in a hypoxic sample. Lane 1: CoCl2 treated Cos-7 nuclear extract (hypoxic). Lane 2: Untreated Cos-7 nuclear extract (normoxic).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of mouse HIF-1 alpha on hypoxia treated MEFs

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody [NB100-449] - Samples: Whole cell lysate (5, 15 and 50 ug) from HeLa cells that were treated with cobalt chloride (+; 200 uM) or mock treated (-). Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody NB100-449 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with exposure times of 30 seconds.

Western Blot: HIF-1 alpha Antibody [NB100-449] - BMDM were seeded at 0.5x10^6 overnight. Cells were treated with 10 ng/ml LPS for 24 hrs. Image from verified customer review.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of total protein using anti-HIF-1 alpha, -CAIX, -ISCU antibodies and VDACs antibody (PMID: 29596470).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of total protein using anti-HIF-1 alpha, -ISCU, -FXN antibodies. CoCl2 was used to treat or not HeLa cells for 2 days (PMID: 29596470).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1 alpha by western blot. Samples: Whole cell lysate (15 and 50 ug) from HEK293T cells that were either treated with 200 uM cobalt chloride (+) or mock treated (-). Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody NB100-449 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with exposure times of 3 minutes.

Western Blot: HIF-1 alpha Antibody [NB100-449] - 3T3-L1 mouse embryonic fibroblast adipose-like cell line. Antibody at 1:2000. WB image submitted by a verified customer review.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Formaldehyde-fixed asynchronous HeLa cells.

Immunoprecipitation: HIF-1 alpha Antibody [NB100-449] - HIF-1 alpha analysis in HEK293 cells. Image from verified customer review.

Immunoprecipitation: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1 alpha by western blot of immunoprecipitates. Samples: Whole cell lysate (1 mg for IP, 20% of IP loaded) from HEK293T cells that were treated with 200 uM cobalt chloride. Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody NB100-449 used for WB at 1 ug/ml and for IP at 6 ug/mg lysate. Detection: Chemiluminescence with exposure times of 3 minutes.

beta -Glucan size affects the requirement for reactive oxygen species (ROS) in IL-1 beta induction.(B) Human mDCs from healthy donors or chronic granulomatous disease (CGD) patients were stimulated with curdlan or glucan-mp for 8 h. HIF-1 alpha and pro-IL-1 beta protein expression measured by immunoblot (n = 2 donors).

Western Blot: Rabbit Polyclonal HIF-1 alpha Antibody - Analysis of HIF-1 alpha antibody on mouse liver nuclear extracts. Image from a verified customer review.

HIF-1 alpha /2 alpha expression in myeloid-specific KO mice targeting the HIF pathway. (A) Images of the colon of wild-type (WT), myeloid-specific Hif-1a KO (hMRP8 Hif-1a KO) or von Hippel Lindau (Vhl) KO (hMRP8 Vhl KO) mice, immunostained for MRP8 (green) & the DNA-binding regions of Hif-1a mRNA (red). Mice were fed with 5% DSS for 4 days prior to immunostaining analyses. Note that there were no MRP8-positive cells that were positive for Hif-1a mRNA in hMRP8 Hif-1a KO (middle column) mice, but we observed many cells that were double positive for MRP8 & Hif-1a mRNA in hMRP8 Vhl KO mice (right column). (B) Images of the colon of hMRP8 Vhl KO mice fed with 5% DSS as in A, immunostained for MRP8 (green) & HIF-1 alpha (red, upper row) or HIF-2 alpha (red, bottom row). DAPI-stained nuclei are shown in blue. White boxes in A & B indicate the regions magnified in the lower or right images, respectively. Yellow arrowheads in A & B indicate cells positive for both markers. Scale bars: 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29967068), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Analysis of microvascular density & HIF-1 alpha activity. Microvascular density of pulmonary tumors was significantly higher in hyperplastic (B) & tumoral lesions (D) of NNK/NTHi treated mice compared to the NNK treated mice (A, & C) detected by CD105 immunostaining (10X, scale bar = 100 μm). HIF-1 alpha immunostaining after NNK/NTHi combined treatment showed hot-spots of high stromal expression in tumors (F), & high expression in perivascular-peribronchiolar lymphocytes (H). In contrast, low, homogenous expression of HIF-1 alpha was detected in the tumors (E) & perivascular-peribronchiolar lymphocytes (G) of NNK treated mice (40X, scale bar = 25 μm). Image collected & cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-11-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Chronic ethanol feeding enhances tumorigenesis markers during AOM/DSS-induced tumorigenesis. AOM/DSS-induced colonic tumorigenesis was induced with (EF) or without (PF) 4 % ethanol feeding as described in Methods section. Cryosections of distal colon were stained for VEGF (a) pSmad (b), or HIF1 alpha (c), & co-stained for F-actin & nucleus. Images presented are representative of n of 5 for each group Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26951793), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB100-449] - Treatment with HBOT or PFD decreased HIF-1 alpha expression in the AD lesions.IHC for HIF-1 alpha (A). Scale bar is 100 µm. HIF-1 alpha + area was measured in five fields chosen at random by using image analysis software (B). RT-PCR for HIF-1 alpha (C). NT, not treated. Data are expressed as the mean ± SD (n = 12). *P<0.05, compared with the control group; ¶P<0.05, compared with the AD-NT group; §, P<0.05, compared with the AD-Steroid group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25275529), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB100-449] - Elevated PIK3/AKT/HIF-1 alpha pathways in KL SqCC tumours.(a,b) Analysis of TCGA gene expression (normalized TPM) & PIK3CA (a) & PTEN (b) genomic copy number alteration profiles. Each dot represents one SqCC patient (n=501). Boxes represent the interquartile range & whiskers are drawn to the minimum & maximum. Kruskal–Wallis non-parametric ANOVA, ****P<0.0001, ***P<0.001, **P<0.01. (c) IHC analysis (top) & quantification (bottom) of p63, p-AKT, p-S6, p-4EBP1, HIF-1 alpha & GLUT1 in KL tumours (n=6 each group). Two-tailed t-test, ****P<0.0001, ***P<0.001, **P<0.01. Scale bar, 50 μm. (d) Immunoblot analysis of HIF-1 alpha & GLUT1 in control shGFP & shHIF-1 alpha knockdown SqCC cell lines, HCC95 & HCC1588. All error bars represent the mean±s.e.m. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28548087), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Immunolocalization of HIF1A in limbs.Immunoreactivity was detected in control limbs in the apical ectodermal ridge & interdigital regions (I.R.). The mesenchymal condensations/developing cartilaginous anlagen (Digits) were not immunoreactive. 4-OOHCPA treatment resulted in a concentration- & time-dependent increase in HIF1A immunoreactivity in this area in the 3 & 6 h exposure groups; staining was diminished by 24 h. Four separate replicates were done. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0051937), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Effects of cell stress on VEGF-A & HIF1 alpha expression in Müller cells & Y79 photoreceptors. (A) VEGF-A was measured by ELISA using conditioned media collected from MIO-M1 & Y79 cells, n = 4–5/group in Müller cells & n = 8/group in Y79 photoreceptors, ** p < 0.01; (B) Western blots for HIF1 alpha using cellular proteins, n = 4/group, ** p < 0.01. For MIO-M1 Müller cells: Grey bars: 5 mM glucose. Black bars: 25 mM glucose. For Y79 photoreceptors: Grey bars: 11 mM glucose. Black bars: 25 mM glucose. NS, not significant; LG, low glucose; HG, high glucose. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/18/3/533), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Phenylhydrazine administration results in hypoxia in retinal vessels & in HIF-1 alpha & HIF-2 alpha stabilization in peripheral blood leukocytes.(A) 3-dimensional reconstructed imaging of the superficial vascular plexus of flat mounted retinae of mice (steady state, treated with PHZ & treated with PHZ & peak EIU induction) stained with DAPI, hypoxyprobe & Isolectin B4. (B) Representative analysis of HIFs by western blot: 50ug of protein from total cell lysate of blood leukocytes isolated from mice treated with PHZ compared to untreated; (C) relative densitometry quantification of HIF western blots from PBMC showing mean, n = 3, error bar showing SD, unpaired t test, ***P < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28112274), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Frozen: HIF-1 alpha Antibody [NB100-449] - Ethanol elevates tumorigenic markers & nuclear localization of HIF1 alpha in colonic crypts during AOM/DSS-induced tumorigenesis: AOM/DSS-induced colonic tumorigenesis was induced with or without 4 % ethanol feeding as described in Methods section. a & b Colonic mucosal extracts were immunoblotted for pSmad & VEGF (a), & the band density evaluated using Image J software (b). Values are mean ± SE (n = 3). Asterisks indicate the values that are significantly different (p < 0.05) from corresponding values for AOM/DSS group. c Cryosections of colon were stained for HIF1 alpha (red) & nucleus (blue). Sections of the images from Fig. 2c are enlarged to show the detailed localization of HIF1 alpha in the nucleus. While arrows indicate nuclear co-localization of HIF1 alpha Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26951793), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Dectin-1-induced NADPH oxidase-derived ROS is required for augmented IL-1 beta during TSLPR inhibition. (A) Human mDC derived from healthy donors (HD) or CGD patients were stimulated with SC glucan for 8 h (n = 2 representative donors presented, three separate experiments performed). Pro-IL-1 beta , IL-1 beta , HIF-1 alpha , phospho-AMPK, AMPK & beta -actin were measured by immunoblot. (B–E) Densitometry of cumulative data was performed using Image Studio Lite software with pro-IL-1 beta , IL-1 beta & HIF-1 alpha normalized to beta -actin & phospho-AMPK normalized to total AMPK. Data is reported as percentage of maximal signal observed within each donor (n = 3 independent donors, presented as pooled data). (F–H) Human mDC derived from HD or CGD patients were stimulated with SC glucan, CA glucan or heat killed C. albicans hyphae with anti-TSLPR or IgG isotype control antibodies for 24 h (n = 3 independent donors, presented as pooled data). IL-1 beta was measured in 24-h cell culture supernatants by ELISA. Cumulative data displayed as mean +SEM. Statistical analysis calculated using one-way ANOVA with Bonferroni post-tests (***p = 0.001,*p = 0.05). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31139177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Reactive oxygen species production & expression of stabilized HIF1 alpha & AMPK subunit alpha 2 & in H1299 & H1299r cells(A) H1299, H1299r, P31 & P31r cell were seeded at a density of 250,000 cells/well & left for 24 h. The cells were then stained with 10 μmol/L dichloro-dihydro-fluorescein diacetate (DCFH-DA). Fluorescence was analysed for 10,000 events. Whole cell lysates were prepared from confluent cultures of H1299, H1299r cells. Proteins (90 μg) were resolved in 10% SDS-PAGE gels & transferred to a PVDF membrane. (B) Representative blot of HIF1 alpha protein expression. (C) Representative blot of AMPK subunit alpha 2 protein expression from four independent experiments. (D) Bar chart of AMPK subunit alpha 2 mean for four separate measurements. Statistical analyses was carried out using the student t-test. ** = p<0.01, * = p<0.05. 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: HIF-1 alpha Antibody [NB100-449] - Pyruvate kinase isoform M2 (PKM2) & hypoxia-inducible factor 1 alpha (Hif-1 alpha ) bind directly to the promoter region of PDL1 in primary murine BMDM cells. BMDM cells were treated with TEPP-46 at 50 µM for 1 h prior to stimulation with LPS (100 ng/ml, 24 h) (A). Binding of PKM2 (left) or Hif-1 alpha (right) to HRE1 or HRE4 of the PD-L1 promoter was detected by incubating cell lysates with biotinylated oligonucleotides spanning the relevant HRE promoter region. Protein–oligonucleotide complexes were isolated using streptavidin agarose beads, & proteins were detected by western blotting. Representative of n = 3. Chromatin immunoprecipitation (ChIP)-PCR (B) using PKM2 & HIF-1 alpha antibodies & primers specific for three promoter regions of PD-L1 (HRE1, 2–3, & 4) & a known Hif-1 alpha binding region of vascular endothelial growth factor as a positive control showing binding of Hif-1 alpha (left) & PKM2 (right) to the PD-L1 promoter in LPS-treated BMDMs (100 ng/ml, 24 h). (C) Sequential ChIP assays measuring simultaneous endogenous binding of PKM2 & Hif-1 alpha to chromatin in response to LPS (100 ng/ml, 24 h) ±TEPP-46 (pretreatment using 50 µM, 60 min). ChIP data are calculated as percent of input, error bar represents mean ± SEM, & statistics are performed as two-tailed unpaired t-test *P < 0.05, **P < 0.01, & ***P < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29081778), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Changes in Hepatic Oxygen Saturation & Hypoxia-related Protein Expression. (A) In the representative case, the hepatic oxygen saturation in the experimental group was lower than that in the control group at four, six, & eight weeks (51.6 vs. 54.5%, 48.4 vs. 53.9%, 32.3 vs. 43.2%, & 31.2 vs. 42.9%). (B) There was significant decrease of oxygen saturation of liver parenchyma in the experimental group compared to those of control group (45.2 vs. 51.9%, 43.0 vs. 49.0%, & 31.3 vs. 49.7%) at the four, six, & eight weeks, respectively (P < 0.001). (C) In the Western blot analysis, the experimental group was relatively higher expression of hypoxia-inducible factor-1 alpha (HIF-1 alpha ) & vascular endothelial growth factor (VEGF) protein than the control group at eight weeks. The full-length blots with these antibodies were presented in supplementary Figure S1. (D,E) The experimental group showed significantly higher expression of HIF-1 alpha & VEGF protein than the control group at eight-week (1.47 ± 0.48 vs. 0.19 ± 0.08 & 0.96 ± 0.23 vs. 0.33 ± 0.06, P = 0.019 & P < 0.018). *P < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29079853), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - BCAT1 expression & inhibition in human macrophages.(a) RNA-sequencing in human monocyte-derived macrophages (hMDMs) shows mRNA copies of BCAT enzymes in normalized counts. n=14 healthy donor hMDMs were used for RNA-seq. (b) Chemical structure of ERG240 & leucine. (c) Enzymatic activity of recombinant human BCAT1 & BCAT2 in presence of ERG240. (d) Relative expression of IRG1, HIF1A & IL1B measured by qRT-PCR in control, ERG240-treated (20 mM, 3 h), LPS treated (100 ng ml−1, 3 h), & LPS+ERG240 treated (LPS, 100 ng ml−1; ERG240, 20 mM for 3 h) hMDMs. ns, non-significant. All expression values are normalized to those obtained for HPRT gene expression. n=3 healthy donor hMDMs were used in each group. (e) Western blot analysis of IRG1, IL-1 beta , HIF-1 alpha & beta-actin (ACTB) in control (untreated), ERG240-treated (20 mM, 3 h), LPS treated (100 ng ml−1, 3 h) & LPS+ERG240 treated (LPS, 100 ng ml−1; ERG240, 20 mM for 3 h) hMDMs. The experiment is representative of three independent experiments using n=3 healthy donor hMDMs each. (f) GC/MS results showing itaconic acid production in hMDMs in presence or absence of ERG240 (20 mM). LPS treatment was for 8 h (100 ng ml−1). GC/MS plot for each donor is shown separately where itaconate abundance denotes arbitrary units. Error bars are s.e.m. Significance was tested using two-tailed Student’s t-test or one-way ANOVA. * P<10−3 following two-way ANOVA. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms16040), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - HIF-1 alpha /2 alpha expression in myeloid-specific KO mice targeting the HIF pathway. (A) Images of the colon of wild-type (WT), myeloid-specific Hif-1a KO (hMRP8 Hif-1a KO) or von Hippel Lindau (Vhl) KO (hMRP8 Vhl KO) mice, immunostained for MRP8 (green) & the DNA-binding regions of Hif-1a mRNA (red). Mice were fed with 5% DSS for 4 days prior to immunostaining analyses. Note that there were no MRP8-positive cells that were positive for Hif-1a mRNA in hMRP8 Hif-1a KO (middle column) mice, but we observed many cells that were double positive for MRP8 & Hif-1a mRNA in hMRP8 Vhl KO mice (right column). (B) Images of the colon of hMRP8 Vhl KO mice fed with 5% DSS as in A, immunostained for MRP8 (green) & HIF-1 alpha (red, upper row) or HIF-2 alpha (red, bottom row). DAPI-stained nuclei are shown in blue. White boxes in A & B indicate the regions magnified in the lower or right images, respectively. Yellow arrowheads in A & B indicate cells positive for both markers. Scale bars: 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29967068), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Increased infiltration of myeloid cells expressing HIF-1 alpha in DSS-induced colitis. (A) Western blot analysis of MRP8, F4/80 or actin using the whole colon tissue lysate from mice fed with water or 5% DSS for 4 days. (B) Images of the colon of mice fed with water or 5% DSS, immunostained for myeloid cells using anti-MRP8 (green) or anti-HIF-1 alpha (red) antibodies. Nuclei were counterstained with DAPI. White rectangles indicate the areas magnified in the images shown below. Yellow arrowheads indicate cells with colocalization of MRP8 & HIF-1 alpha . Quantification of MRP8-positive cells & percentage of those expressing HIF-1 alpha are shown as bar graphs on the right-hand side. Data are mean±s.e.m. for at least three independent fields examined per mouse (n≥3 per group). **P<0.01 & ***P<0.001, assessed by Student's t-test. HPF, high-powered field. (C) Images of the colon of mice fed with 5% DSS, immunostained for MRP8 (red) & CD11b (green). DAPI-stained nuclei are shown in blue & a merged image is also shown. Scale bars: 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29967068), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Increased HIF-1 alpha expression is regulated by TSLPR inhibition & not IL-1 beta secretion (A) Human mDC were stimulated with SC glucan with either anti-TSLPR or IgG isotype control antibodies for 8 h in the presence or absence of IL-1 beta neutralization antibodies (n = 1 representative donor presented, three separate experiments performed). Pro-IL-1 beta , IL-1 beta , HIF-1 alpha , phospho-p38 MAPK, p38 MAPK, phospho-AMPK, AMPK & beta -actin were measured by immunoblot. (B,C) Densitometry of cumulative data was performed using Image Studio Lite software with HIF-1 alpha normalized to beta -actin & phospho-AMPK normalized to AMPK. Data is reported as percentage of maximal signal observed within each donor (n = 3 independent donors, presented as pooled data). Cumulative data displayed as mean +SEM. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31139177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - miR-18b (miR-18b-5p) regulates Hif1 alpha & reduces apoptosis in mtNSC-34 cells. a Overexpressed miR-18b (miR-18b-5p) decreased Hif1 alpha & Mef2c proteins. Both Mctp1 & Rarb expression were increased by miR-18b (miR-18b-5p). Downregulated Bax & upregulated Bcl2 by miR-18b (miR-18b-5p) diminished apoptosis in mtNSC-34 cells. b RT-qPCR analysis showed low expression of Hif1 alpha & Mef2c mRNAs. c Mctp1 & Rarb transcripts were highly expressed by miR-18b (miR-18b-5p). d Bax mRNAs were decreased & Bcl2 mRNAs were increased by overexpressed miR-18b. e miR-18b (miR-18b-5p) was overexpressed in mtNSC-34 cells. f miR-206 was reduced by miR-18b (miR-18b-5p). g LDH release analysis explained that transfected miR-18b (miR-18b-5p) restores apoptosis. h Luciferases assay with 3′ UTR of Hif1 alpha showed that Hif1 alpha is target of miR-18b in contNSC-34 cells. i & j Overexpression of miR-18b (miR-18b-5p) enhanced neuronal differentiation (MAP2) & attenuated intracellular Ca2+ levels (Cont (0.098) versus miR-18b (miR-18b-5p) (0.051) in fluorescence intensities from baseline 490/525 ratio) in mtNSC-34 cells. Empty vector served as a negative control (Cont). Arrow represents SOD1 aggregation (green). Scale bar, 40 μm. Significantly different at *, p < 0.05; **, p < 0.005. The experiments were replicated 5 times Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32605607), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Hypoxic conditions upregulate CNN3 in BeWo cells.BeWo cells were cultured either under normoxic or under hypoxic conditions for 24(A) Total protein lysates were examined with the Western Blot technique to detect CNN3 protein levels. For normalization, HPRT was stained on the same membrane. As hypoxia marker, HIF-1 alpha was detected as well. (B) Protein bands of CNN3 & HPRT were densitometrically measured on the Western Blot membrane & CNN3/HPRT levels are plotted in the graph (white column: normoxia; black column: hypoxia). n = 3. (C) BeWo cells were serum starved for 16 h & then treated with either PBS as control or 200 µM CoCl2 for 6 h in serum free medium. Then total protein was isolated & a Western Blot was performed. The HIF-1 alpha, CNN3 & HPRT protein was detected on the membrane with specific antibodies. (D) A densitometric analysis was performed to determine CNN3/HPRT levels (white column: normoxia; black column: hypoxia). n = 3. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25050546), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Dynamics of the hypoxia-triggered decrease in MET phosphorylation & its reversal upon reoxygenation(A) MCF10A cells were incubated under normoxia or hypoxia for 1 h. They were then treated, under the same oxygen pressure, with 10 ng/mL HGF/SF for 5, 10, 20, 30, 60, 90 & 120 minutes. A control (Ctrl) without any HGF/SF stimulation was also performed. The same amount of protein was analyzed by western blotting with antibodies directed against: phosphorylated residues in the MET kinase domain, the MET kinase domain, the hypoxia marker HIF1a, phosphorylated Akt, Akt, Erk2, phosphorylated Erk, & actine. The positions of prestained molecular weight markers are indicated. Arrows indicate the positions of precursor & mature full-length MET. (B) MCF10A cells were incubated under hypoxia for 5, 10, 15 or 30 minutes. Another set of cells were incubated under hypoxia for 1 h & then returned to normoxia for 5, 10, 15 or 30 min (re-oxygenation). A control under normoxic (N) conditions was also included. The same amount of protein was analyzed by western blotting as previously described with the addition of GAB1 & its phosphorylated form. (C) MCF10A cells were placed under normoxic or hypoxic conditions for 1 h or hypoxia for 1 h & then normoxia for 10 minutes (reoxygenation). Cells were then treated at the indicated time with 10 ng/mL of HGF/SF. Cell lysates were incubated for AlphaScreen specific phospho-Erk & phospho-Akt quantitation. Error bars represent standard deviations (n = 3; ± SD). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29930749), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - The role of HIF-1 in purinosome formation.a, quantifying the number of purinosome-containing cells in normoxia or hypoxia (24 h) in purine-rich medium & normoxia in purine-depleted medium (purine -ve), cells in hypoxia transfected with siRNA to HIF-1 alpha (+ siRNA), & cells in purine-rich medium supplemented with DFX. Data shown are n = 3, mean ± S.E., total number of cells counted are shown in parentheses. b, time course of purinosome formation in hypoxia shows the number of purinosome-containing cells steadily increases after 3 h in hypoxia. Re-oxygenation of the samples after hypoxic incubation for 10 h reverts the number purinosome-containing cells back to normoxic levels. Data shown is n = 3, mean ± S.E., total number of cells counted are shown in parentheses. c, time course of HIF-1 alpha stabilization in hypoxia shows maximum HIF-1 alpha protein expression levels at 3 h in hypoxia, after which the HIF-1 alpha expression decreases. The positions of molecular markers are shown for each blot; uncropped blots with overlaid markers are deposited in the raw data files. d, the effect of hypoxia on the transcription of purine biosynthesis enzymes measured by qPCR. Vascular endothelial growth factor (VEGF) & HIF-1 alpha are controls. Data shown are n = 5, mean ± S.E. e, the effect of hypoxia on the protein expression levels of the purine biosynthetic enzymes. HIF-1 alpha is stabilized in hypoxia as expected, & no significant increase in the purine enzymes was detected between normoxic (21% oxygen) & hypoxic (1% oxygen) growth conditions. The positions of molecular markers surrounding each band of interest are shown for each blot; uncropped blots with overlaid markers are deposited in the raw data files. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32439803), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - C/EBPδ promotes HIF-1 alpha expression in macrophages through inhibition of FBXW7 alpha a) RT-PCR analysis of FBXW7 isoform expression from different sources as follows. 1, primary peritoneal macrophages (PPMs); 2, RAW 264.7 cells; 3, MMTV-Neu mammary tumour tissue; 4, primary mouse embryo fibroblasts. Numbers indicate the position of size markers in base pairs. (b) RT-qPCR analysis of Fbxw7 transcript levels in PPMs from WT & Cebpd−/− KO mice, cultured +/− LPS (100 ng/ml, 24 h), compared to WT untreated (n=4, *P<0.05; **P<0.001). (c) Western analysis of nuclear extract (NE) from primary human monocytes nucleofected with siRNA oligos (C, control; D, CEBPD; F, FBXW7) & treated with LPS (100 ng/ml) & 1%O2 (16 h) as indicated. SE, short exposure; LE, long exposure. (d) RT-qPCR analysis of FBXW7 & CEBPD transcripts in primary human monocytes as in panel (c) (n=3, *P<0.05; **P<0.001). (e) Western analysis of NE from PPMs nucelofected with siRNA oligos & treated with LPS (100 ng/ml) & 1%O2 for 16 h as indicated. SE, short exposure; LE, long exposure.Where applicable, data are mean±S.E.M., evaluated by two-tailed unequal variance t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23575666), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Hypoxia downregulates CFTR expression. (a) Phase-contrast photographs of MDCK cells treated with normoxia & hypoxia conditions; (b) Western blot showing downregulation of CFTR in the MDCK cells by hypoxia (Full-length blot is shown in Supplementary Figure S7b.); (c) Real time-PCR assay showing decreased mRNA expression of CFTR induced by hypoxia in HK-2 cells,*p < 0.05; (d) Western blot showing decreased expression of CFTR induced by hypoxia in HK-2 cells (Full-length blot is shown in Supplementary Figure S7c.); (e) Western blot showing the expression changes of HIF-1 alpha & EMT markers induced by hypoxia in MDCK cells, quantification analysis is shown in the lower panel, *p < 0.05; (Full-length blot is shown in Supplementary Figure S7d.) (f) Immunofluorescent staining showing dramatically increased HIF-1 alpha & reduced CFTR protein levels in tubular epithelial cells at inner cortices in UUO kidney (arrow). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28701694), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - NRP1 & MET on HT1080 cells are accessible for blood-borne rhodocetin-alpha beta (A) expression of NRP1 & MET by HT1080 cells was proven by flow cytometry. Gray, isotype-matched controls. (B) flow cytometry of NRP1-knockout HT1080 cells demonstrating their NRP1-deficiency & unaffected MET expression. (C) treatment with CoCl2, mimicking a hypoxic tumor micro-environment, induced upregulation of HIF-1 alpha . (D) NRP1, as a downstream target of HIF-1 alpha , is upregulated in HT1080 cells but not in NRP1-knockout HT1080 cells. beta -actin immunoblots show even loading. (E) increased HIF-1 alpha (red) levels in hypoxic tumor regions, which also contained partly (arrows) or completely (open arrows) EC-deficient VM vessels. ECs are stained green & nuclei blue. (F) immunostaining of NRP1 (red) & MET (blue) showed that both proteins were present on HT1080 cells & ECs in tumor tissue. Note the continuity between EC-lined vasculature & EC marker-deficient vessels (arrows in F’). F’, F’’, in oblique view, gating of the green CD31 signal also showed an apical absence of NRP1 on ECs in contrast to MET (open arrows). (G) the fluorescence intensity along a traceroute, averaged over a width of 5 pixels, (rectangle in F) through the endothelium revealed that in ECs NRP1, unlike MET, is absent from the apical side & restricted to the basolateral side. In contrast, on ATV/VM-lining cells (F’, arrows) both NRP1 & MET are accessible from the bloodstream (H). Vertical gray lines in G & H indicate the position of the apical cell border. Original magnification was 400× (E) & 630× (F-F’’). Representative images are shown. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.25032), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - NRP1 & MET on HT1080 cells are accessible for blood-borne rhodocetin-alpha beta (A) expression of NRP1 & MET by HT1080 cells was proven by flow cytometry. Gray, isotype-matched controls. (B) flow cytometry of NRP1-knockout HT1080 cells demonstrating their NRP1-deficiency & unaffected MET expression. (C) treatment with CoCl2, mimicking a hypoxic tumor micro-environment, induced upregulation of HIF-1 alpha . (D) NRP1, as a downstream target of HIF-1 alpha , is upregulated in HT1080 cells but not in NRP1-knockout HT1080 cells. beta -actin immunoblots show even loading. (E) increased HIF-1 alpha (red) levels in hypoxic tumor regions, which also contained partly (arrows) or completely (open arrows) EC-deficient VM vessels. ECs are stained green & nuclei blue. (F) immunostaining of NRP1 (red) & MET (blue) showed that both proteins were present on HT1080 cells & ECs in tumor tissue. Note the continuity between EC-lined vasculature & EC marker-deficient vessels (arrows in F’). F’, F’’, in oblique view, gating of the green CD31 signal also showed an apical absence of NRP1 on ECs in contrast to MET (open arrows). (G) the fluorescence intensity along a traceroute, averaged over a width of 5 pixels, (rectangle in F) through the endothelium revealed that in ECs NRP1, unlike MET, is absent from the apical side & restricted to the basolateral side. In contrast, on ATV/VM-lining cells (F’, arrows) both NRP1 & MET are accessible from the bloodstream (H). Vertical gray lines in G & H indicate the position of the apical cell border. Original magnification was 400× (E) & 630× (F-F’’). Representative images are shown. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.25032), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Dectin-1-induced TSLP negatively regulates pro-IL-1 beta & HIF-1 alpha . (A) Human mDC were stimulated with SC glucan, CA glucan or heat killed C. albicans hyphae with anti-TSLPR antibodies or IgG isotype control for 24 h (n = 6 independent donors, presented as pooled data). Lactate production was measured in cell-culture supernatants using colourmetric L-lactate detection kit. (B) Human mDC were stimulated SC glucan with either anti-TSLP, anti-TSLPR or IgG isotype control antibodies for 8 h (n = 1 representative donor presented, three separate experiments performed). Pro-IL-1 beta , IL-1 beta , HIF-1 alpha , phospho-p38 MAPK, p38 MAPK, phospho-AMPK, AMPK & beta -actin were measured by immunoblot. (C–G) Densitometry of cumulative data was performed using Image Studio Lite software with pro-IL-1 beta , IL-1 beta & HIF-1 alpha normalized to beta -actin & phospho-p38 MAPK & phospho-AMPK normalized to total p38 MAPK & AMPK respectively. Data is reported as percentage of maximal signal observed within each donor (n = 3 independent donors, presented as pooled data). Cumulative data displayed as mean +SEM. Statistical analysis calculated using one-way ANOVA with Bonferroni post-tests (***p = 0.001). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31139177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Involvement of MET in Akt & Erk pathway activation under hypoxiaMCF10A cells were transfected with a pool of three MET-targeting siRNAs (20 nM) or a control siRNA (siCtrl). A control without siRNA was also included (Ctrl) (A). MCF10A cells were transfected with two HIF1a-targeting siRNAs (20 nM), independently or together, or a control siRNA (siCtrl) (B). The cells were then placed for 1 h under normoxic or hypoxic conditions & treated or not for 10 min with 10 ng/mL HGF/SF. In each experiment, the same amount of protein was analyzed by western blotting with antibodies directed against: phosphorylated residues in the MET kinase domain, the MET kinase domain, phosphorylated Akt, Akt, phosphorylated Erk, Erk2, phosphorylated GAB1, GAB1, or hypoxia marker HIF1a. The positions of prestained molecular weight markers are indicated. Arrows indicate the positions of precursor & mature full-length MET & Erk1/2 proteins. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29930749), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - beta -Glucan size affects the requirement for reactive oxygen species (ROS) in IL-1 beta induction. (A) Human monocyte-derived dendritic cell (mDC) stimulated with curdlan or glucan-mp for 8 h. ROS were detected by incubating cells with CellRox Green fluorescence dye & analysis by flow cytometry (representative experiment of two) presented, (B) Human mDCs from healthy donors or chronic granulomatous disease (CGD) patients were stimulated with curdlan or glucan-mp for 8 h. HIF-1 alpha & pro-IL-1 beta protein expression measured by immunoblot (n = 2 donors). (C–L) Human mDC from healthy donors or CGD patients were stimulated with curdlan or glucan-mp for 24 h (n = 6 donors). IL-1 beta , IL-6, IL-23, TSLP, & CCL22 secretion were measured by enzyme-linked immunosorbent assay, with cumulative data displayed as mean ± SEM. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fimmu.2017.00791/full), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ca, Ch, Gt, Pm, Pm, Bv

Species Glossary

Reactivity: Hu, Mu, Rt, Ca, Ch, Gt, Pm, Pm, Bv

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Validated by:

Validated by:

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NB100-2331

LC3A Antibody - BSA Free

(22 Reviews)

(309 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 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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NB600-308

GFP Antibody - BSA Free

(18 Reviews)

(307 Publications)

0.1 mg

NB600-308

Analysis of GFP in transgenic mouse pancreas in OCT. The blue region in the center is an islet which does not express GFP and is negative. Image from verified customer review.

Analysis of GFP tagged PKA subunits over-expressed in HEK293T cells using GFP antibody. Image from verified customer review.

Staining of GFP+ hepatocytes (brown) transplanted into the liver of a recipient animal using anti-GFP antibody. Cyclin D1 double staining in blue. Image from verified customer review.

Total cell lysates (20ug) with protein of interest tagged with GFP was loaded. Blotted with GFP antibody [HRP] NB600-308H, without any secondary antibody. Image from verified customer review.

Germaria from wt or chico1/KG mutant ovaries endogenously expressing Myc-GFP stained with anti-GFP, anti-1B1, and DAPI. Scale bars, 10 um. (F) Quantification of relative Myc-GFP intensity in germarium region 2B from wt or chico1/KG mutant ovaries. n = 10 germaria for each genotype. Error bars represent SEM. *p<0.005. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31612862/) licensed under a CC-BY license.

Western Blot of GFP antibody. Lane 1: Wild type GFP (0.1 ug) was used to spike HeLa whole cell lysate. Lane 2: none. Load: 30 ug per lane.Primary antibody: GFP antibody at 1:1000 for overnight at 4C.Secondary antibody: IRDye800(TM) Goat-a-Rabbit IgG [H&L] MX10 at 1:10,000 for 45 min at RT.Block: 5% BLOTTO in PBS overnight at 4C.Predicted/Observed size: 27 kDa for epitope tag GFP. Other band(s): none.

Western Blot of GFP antibody. Lane 1: 293FT cells transfected with CDK4 dominant negative (C-).Lane 2: 293FT cells positive control (C+).Load: 25 ug per lane.Primary antibody: GFP antibody at 1:400 for overnight at 4C.Secondary antibody: IRDye800(TM) rabbit secondary antibody at 1:10,000 for 45 min at RT.Block: 5% BLOTTO overnight at 4C.Predicted/Observed size: 27 kDa for GFP.

Western Blot of GFP antibody. Marker: Opal Pre-stained ladder. Lane 1: HEK293 lysate. Lane 2: HeLa Lysate. Lane 3: CHO/K1 Lysate. Lane 4: MDA-MB-231. Lane 5: A431 Lysate. Lane 6: Jurkat Lysate. Lane 7: NIH/3T3 Lysate. Lane 8: E-coli HCP Control. Lane 9: FLAG Positive Control Lysate. Lane 10: Red Fluorescent Protein. Lane 11: Green Fluorescent Protein. Lane 12: Glutathione-S-Transferase Protein. Lane 13: Maltose Binding Protein. Load: 10 ug of lysate or 50ng of purified protein per lane. Primary antibody: GFP antibody at 1ug/mL overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:30,000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS for 30 min at RT.Predicted/Observed size: 30 kDa for GFP.

Adult mouse brain tissue. This image was submitted via customer review.

Immuno-microscopy of GFP antibody. Monocyte derived dendritic cells and dermal macrophages were challenged and directly visualized with eGFP labeled Dengue virus to localize sequestration of virus particles in the different cells (upper). The location of the GFP was confirmed by TEM (lower magnified view) using rabbit anti GFP Primary antibody (1:200) and a gold labeled secondary antibody.

Analysis of GFP in mouse retinal cells. Image courtesy of anonymous customer review.

IP analysis of GFP in S2 Cells (Drosophila). Image from verified customer review.

Immunoprecipitation experiments showing that GFP-MISP binds endogenous IQGAP1 (upper panel) and MISP binds to IQGAP1 endogenously using specific antibodies (lower panel) in mitotically arrested (taxol and nocodazole, respectively) HeLa cell lysates. Image collected and cropped by CiteAb from the following publication (nature.com/articles/s41598-018-24682-8), licensed under a CC-BY license.

Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB600-308] - Myc is post-transcriptionally regulated by the IIS-GSK3 cascade.(A) Germaria from ovaries of ctrl, chicoRNAi, & bskRNAi driven by nos-Gal4 expressing LacZ under the control of myc endogenous promoter stained with anti-beta -galactosidase & anti-1B1. Scale bars, 10 μm. (B) A representative live imaging of germaria from flies expressing GFP-tagged Sgg under its endogenous promoter. Scale bar, 10 μm. (C) A germarium from wt flies endogenously expressing Myc-GFP stained with anti-GFP (green) & anti-p-GSK3 (red). Germarium regions are indicated. Myc protein highly corresponds to inhibited form of GSK3. Scale bar, 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31612862), licensed under a CC0-1.0 license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB600-308] - Myc protein pattern in the ovary.(A) A representative low-magnification image of ovarioles from flies endogenously expressing Myc-GFP stained with anti-GFP, anti-1B1, & DAPI. Note that Myc level is low in early germarium stages, becomes high from germarium region 2B, & reduces from the stage-10 egg chamber. Scale bar, 50 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31612862), licensed under a CC0-1.0 license. Not internally tested by Novus Biologicals.

Western Blot: GFP Antibody [NB600-308] - MISP interacts & co-localizes with IQGAP1. (a) HEK293T cells transiently overexpressing GFP-IQGAP1 & FLAG-MISP were used for co-immunoprecipitation experiments from both sides (GFP trap or FLAG M2 beads). (b) Endogenous immunoprecipitation using IQGAP1 antibody or control immunoglobulin (IgG) was carried out in HeLa cell lysates & MISP was detected in the eluate. (c) In vitro interaction between purified MBP-MISP & GST-IQGAP1 was detected by GST & MBP pull-down experiments. (d) Immunoprecipiation experiments showing that GFP-MISP binds endogenous IQGAP1 (upper panel) & MISP binds to IQGAP1 endogenously using specific antibodies (lower panel) in mitotically arrested (taxol & nocodazole, respectively) HeLa cell lysates. (a–d) Images were gained from the same Western blot membrane after cutting/cropping & presented with different exposure times or contrast enhancement for better presentation purposes. The dividing lane marks the grouping of images of the same (or different) membrane. Uncropped blots, where applicable, are included in Supplementary Fig. 4. (e) HeLa cells were immunostained for MISP & IQGAP1 & co-localization was visualized in mitosis in single-plane confocal images, scale bar: 5 μm. Lower pictures: After manual thresholding white spots mark the co-localizing areas in the merge images. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29679050), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB600-308] - BRC-1 & BRD-1 are inter-dependent for localization.A) Co-localization between BRD-1::GFP (green) & TagRFP-T::BRC-1 (red) at late pachytene in live worms. Scale bar = 10 μm. B) Stills of germline nuclei from live worms expressing GFP::BRC-1 & mCherry::Histone H2B (mCherry::his-58; gfp::brc-1); merge & GFP channel; top two panels, respectively. GFP::BRC-1 expression in brd-1(ok1623) mutant at indicated meiotic stages. Bottom two panels show BRD-1::GFP localization in wild type & the brc-1(xoe4) mutant. Images are projections through half of the gonad. TZ = transition zone; EP = early pachytene; MP = mid pachytene; LP = late pachytene; DP = diplotene; DK = diakinesis. Scale bar = 5 μm. C) Immunoblot of whole worm extracts from indicated worms probed with anti-GFP & alpha -tubulin antibodies. Lane 1 = N2: wild type; Lane 2 = JEL515: gfp::brc-1; Lane 3 = JEL520: gfp::brc-1 brd-1(ok1623); Lane 4 = JEL744: brc-1(xoe4) brd-1::gfp; Lane 5 = JEL657: brd-1::gfp; Lane 6 = JEL678: brc-1(tm1145) brd-1::gfp. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30383767), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GFP Antibody [NB600-308] - BRC-1 & BRD-1 are inter-dependent for localization.A) Co-localization between BRD-1::GFP (green) & TagRFP-T::BRC-1 (red) at late pachytene in live worms. Scale bar = 10 μm. B) Stills of germline nuclei from live worms expressing GFP::BRC-1 & mCherry::Histone H2B (mCherry::his-58; gfp::brc-1); merge & GFP channel; top two panels, respectively. GFP::BRC-1 expression in brd-1(ok1623) mutant at indicated meiotic stages. Bottom two panels show BRD-1::GFP localization in wild type & the brc-1(xoe4) mutant. Images are projections through half of the gonad. TZ = transition zone; EP = early pachytene; MP = mid pachytene; LP = late pachytene; DP = diplotene; DK = diakinesis. Scale bar = 5 μm. C) Immunoblot of whole worm extracts from indicated worms probed with anti-GFP & alpha -tubulin antibodies. Lane 1 = N2: wild type; Lane 2 = JEL515: gfp::brc-1; Lane 3 = JEL520: gfp::brc-1 brd-1(ok1623); Lane 4 = JEL744: brc-1(xoe4) brd-1::gfp; Lane 5 = JEL657: brd-1::gfp; Lane 6 = JEL678: brc-1(tm1145) brd-1::gfp. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30383767), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: GFP Antibody [NB600-308] - Other IIS components are not altered by bsk RNAi.(A) Confocal images for germaria expressing Chico-GFP, myc-Dp110 or PDK1-GFP in the background of ctrl or bskRNAi from ovaries stained with anti-GFP or anti-myc. Scale bars, 10 μm. (B) Ovaries expressing LacZ driven by the puc promoter stained with anti-beta -galactosidase, anti-1B1, & DAPI. Note that JNK activity is moderately induced in late germarium & decreased in the following stages, while JNK activity is much stronger in follicle cells of a maturing egg. Scale bars, 20 μm. (C) Germaria from ovaries of ctrl & bskRNAi driven by nos-Gal4 stained with anti-AKT & anti-1B1. Scale bars, 10 μm. (D) Quantification of AKT intensity from cysts in germarium region 2A & 2B of ovaries with ctrl & bskRNAi driven by nos-Gal4. Intensities are normalized to the value of ctrl at region 2B. n = 7 & 8 germaria for ctrl & bskRNAi, respectively. Error bars represent SEM. *p<0.005. (E) Visualization of the InR mRNA by FISH in germaria from ovaries of ctrl & bskRNAi driven by nos-Gal4. Note that InR mRNA level is decreased by bsk RNAi. Germaria are outlined with dotted lines. Scale bars, 10 μm. (F) Quantification of InR mRNA density in region 2A & 2B germ cells from ovaries with indicated genotypes. n = 9 germaria for each genotype. Error bars represent SEM. *p<0.005. (G) Quantification of relative Myc-GFP intensity in region 2B cysts of ovaries illustrated in Figure 5H. n = 10 germaria for each genotype. Error bars represent SEM. *p<0.005.10.7554/eLife.49309.017Figure 5—figure supplement 1—source data 1.Relative AKT intensity, InR mRNA density, & Myc-GFP intensity in the germarium.Relative AKT intensity, InR mRNA density, & Myc-GFP intensity in the germarium. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31612862), licensed under a CC0-1.0 license. Not internally tested by Novus Biologicals.

$Western Blot: GFP Antibody [NB600-308] - GFP Expression within A. thaliana upon infiltration with SWNT-PM-CytKH9/pDNA complexes.a Reporter construct design of transient GFP expression for the nucleus & mitochondria. b Representative confocal laser scanning microscopy images of A. thaliana root cells from a minimum of five seedlings for each condition (n = 5) 18 h post infiltration with SWNT-PM-CytKH9/pDNA complexes containing pDONR-35S-GFP or pDONR-Cox2-GFP reporter constructs for nuclear or mitochondrial expression, respectively. Mitochondria were stained with MitoTracker Red (CMXRos), & colocalization analysis was performed on GFP expression & MitoTracker signals. Scale bars represent 20 μm. c, d Representative western blots & their quantification from cytosolic (c) & mitochondrial (d) fractions isolated from approximately 60 seedlings 18 h post infiltration with SWNT NCs probed for GFP expression. Actin & cytochrome c were used as loading controls for cytosolic & mitochondrial proteins, respectively. Data points from five biological replicates (n = 5) are represented as the mean ± standard deviation. Statistical significance was determined by Brown-Forsythe & Welch one-way ANOVA test. For the cytosolic samples, P-values are 0.0027, & 0.0020 between DNA only & SWNT-PM-CytKH9, SWNT-PM-Cytcox, respectively, & 0.0253 between SWNT-PM-CytKH9 & SWNT-PM-Cytcox. For the mitochondrial samples, P-values are 0.0019, & 0.0333 between DNA only & SWNT-PM-CytKH9, & SWNT-PM-Cytcox, respectively, & 0.0359 between SWNT-PM-CytKH9 & SWNT-PM-Cytcox. ns – not statistically significant, *P < 0.05, **P < 0.01. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35577779), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: GFP Antibody [NB600-308] - VGLL1 regulates MMP9 transcription in gastric cancer cells. (a,b) Viability (a) & invasion (b) assay of NUGC3 cells. Cells were treated with 20 nM siScramble (SC) or siMMP9 for 48 h, & then stained with sulforhodamine B. Data are presented as mean ±SD. n = 3; * p < 0.05 (Student’s t-test). (c) MMP9 mRNA expression regulation by VGLL1 in gastric cancer cells treated with siVGLL1 or siSC (control) was measured by qPCR. (d) Changes in MMP9 expression upon knockdown or overexpression of VGLL1 assayed by western blotting in NUGC3 cells. (e) IHC of VGLL1 & MMP9 expression in the liver & lungs of an in vivo metastasis mouse model using surgical resection of tumors. Scale bar, 200 µm. (f) Construction of various luciferase reporter systems under control of the MMP9 promoter. (g) MMP9 promoter activities of the reporter systems containing modified TEA-binding sites were measured in NUGC3 cells. n = 3; * p < 0.05 (Student’s t-test). (h) Effect of TEAD4 on VGLL1-regulated MMP9 transcriptional activity. NUGC3 cells treated with siSC or siTEAD4 for 24 h were transfected with MMP9-luc, Renilla-luc, pCDNA3.1, & pcDNA3.1-myc-VGLL1 vectors for 48 h. n = 3; * p < 0.05 (Student’s t-test). (i) Interaction between VGLL1 & TEAD4. Lysates of NUGC3 cells that were transfected with pcDNA3.1-myc-VGLL1 & pEGFP-N1-TEAD4 were immunoprecipitated using anti-IgG, anti-GFP, & anti-Myc antibodies. Protein expression was analyzed by immunoblotting. (j) ChIP assays while using nuclear extracts of NUGC3 cells treated with siTEAD4. The ChIP-enriched DNA was subjected to PCR. (k) Target genes of VGLL1 & YAP. NUGC3 cells were transfected with pcDNA3, pcDNA3-myc-VGLL1, or pcDNA3-myc-YAP, & the mRNA expression levels were analyzed by RT-PCR. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31816819), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/Immunofluorescence: Rabbit GFP pAb [NB600-308] - Analysis of GFP in human Hs746T gastric cancer cell line. Image from a verified customer review.

Reactivity:

All-NA

Species Glossary

Reactivity: All-NA

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	NB600-308AF350
Alexa Fluor 405	NB600-308AF405
Alexa Fluor 488	NB600-308AF488
Alexa Fluor 532	NB600-308AF532
Alexa Fluor 594	NB600-308AF594
Alexa Fluor 647	NB600-308AF647
Alexa Fluor 700	NB600-308AF700
Alexa Fluor 750	NB600-308AF750
Allophycocyanin	NB600-308APC
Allophycocyanin/Cy7	NB600-308APCCY7
Biotin	NB600-308B
CoraFluor 1	NB600-308CL1
DyLight 350	NB600-308UV
DyLight 405	NB600-308V
DyLight 488	NB600-308G
DyLight 550	NB600-308R
DyLight 594	NB600-308DL594
DyLight 650	NB600-308C
DyLight 680	NB600-308FR
DyLight 755	NB600-308IR
FITC	NB600-308F
HRP	NB600-308H
Janelia Fluor 525	NB600-308JF525
Janelia Fluor 549	NB600-308JF549
Janelia Fluor 646	NB600-308JF646
Janelia Fluor 669	NB600-308JF669
PE	NB600-308PE
PE/Atto594	NB600-308PEATT594
PE/Cy5.5	NB600-308PECY55
PE/Cy7	NB600-308PECY7
PerCP	NB600-308PCP
mFluor Violet 450 SE	NB600-308MFV450
mFluor Violet 500 SE	NB600-308MFV500
mFluor Violet 610 SE	NB600-308MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-308-1mg
BSA Free	NB600-308SS

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MIF00

Mouse IFN-gamma Quantikine ELISA Kit, 2 Plate

(314 Publications)

1 Kit (1 Plate)

MIF00

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

Applications:

Conjugate	Catalog #	Availability	Size	Price
HRP	PMIF00
HRP	SMIF00

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NB110-53818

ATG5 Antibody - BSA Free

(7 Reviews)

(294 Publications)

0.1 ml

NB110-53818

ATG5 & ATG7 enhanced autophagy & inhibited ER stress in chondrocyte. a Western blotting analysis of LC3, P62, ATG5, ATG7 & ATG5-ATG12 expression after infected with Ad-ATG5, Ad-ATG7 & Ad-ATG5 + Ad-ATG7 in the C28I2 cells. beta -actin is served as an internal control. b Qualitative analysis of ATG5, ATG7, ATG5-ATG12, LC3 & P62. The values were normalized to beta -actin. c C28I2 cells were double stained with LC3 (red) & DAPI (blue) & visualized by confocal microscopy (400X) after treated with Rapamycine, Ad-ATG5, Ad-ATG7 & Ad-ATG5 + Ad-ATG7 24 h. *P UAS‐Atg5‐RNAi flies (Atg5‐RNAi). Atg5 was reduced in Atg5‐RNAi flies (n = 6, one‐way ANOVA, *P < 0.05 vs. control). (D, E) Downregulation of autophagy abolished the beta ‐GPA‐mediated lifespan extension (n = 200, P < 0.0001 vs. control, log‐rank test). For all Western blots, relative band intensity was estimated using IMAGEJ. Data are shown as mean ± SEM. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26120775), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ATG5 Antibody - BSA Free [NB110-53818] - Differences in neuronal autophagy & dendrite varicosity following HIV-1 Tat protein & morphine treatment. (A) Representative images of neurons transfected with a fluorescent reporter plasmid to monitor autophagic flux at 8 h following the indicated treatments. GFP (green) & GFP + mRFP (yellow) fluorescence are observed prior to the fusion of autophagosomes with lysosomes whereas only mRFP (red) fluorescence is present in post-fusion autolysosomes. DIC, differential interference contrast microscopy image. DAPI (blue) staining indicates cell nuclei. (B) Quantification of autolysosomes (red puncta) from (A). F(3,13) = 8.756, p = 0.0019; ∗p < 0.05 when compared to all other groups. (C) Western blotting analysis of the indicated autophagy associated protein levels at 24 h following the indicated treatments. GAPDH was used as a loading control. Blots are representative of three independent experiments. (D) Quantification of dendrite beading from (A). F(3,77) = 6.429, p = 0.0006; ∗p < 0.05 when compared to control cells. Error bars show the SEM. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/Article/10.3389/fmicb.2015.00653/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Al, Bv, Dr, Fi, Gp, Pm, Xp, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Al, Bv, Dr, Fi, Gp, Pm, Xp, Ze

Applications:

, , , , , , , , , , ,

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

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB110-53818AF350
Alexa Fluor 405	NB110-53818AF405
Alexa Fluor 488	NB110-53818AF488
Alexa Fluor 532	NB110-53818AF532
Alexa Fluor 594	NB110-53818AF594
Alexa Fluor 647	NB110-53818AF647
Alexa Fluor 700	NB110-53818AF700
Alexa Fluor 750	NB110-53818AF750
Allophycocyanin	NB110-53818APC
Biotin	NB110-53818B
CoraFluor 1	NB110-53818CL1
DyLight 350	NB110-53818UV
DyLight 405	NB110-53818V
DyLight 488	NB110-53818G
DyLight 550	NB110-53818R
DyLight 594	NB110-53818DL594
DyLight 650	NB110-53818C
DyLight 680	NB110-53818FR
DyLight 755	NB110-53818IR
FITC	NB110-53818F
HRP	NB110-53818H
Janelia Fluor 525	NB110-53818JF525
Janelia Fluor 669	NB110-53818JF669
PE	NB110-53818PE
PerCP	NB110-53818PCP
mFluor Violet 450 SE	NB110-53818MFV450
mFluor Violet 500 SE	NB110-53818MFV500
mFluor Violet 610 SE	NB110-53818MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB110-53818SS

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BC100-494

PINK1 Antibody - BSA Free

(13 Reviews)

(294 Publications)

0.1 ml

BC100-494

$PINK1/Parkin-mediated mitophagy. Western Blot of mitochondrial and cytosolic fractions for PINK1, Parkin, and ubiquitin. E Quantitation of protein levels, normalized to VDAC and GAPDH, Ubiquitin: n = 6, PINK1: n = 5, Parkin: n = 8. R, post-reoxygenation. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33980811/) licensed under a CC-BY license.$

Confocal microscopy analysis of the mitophagy initiation in the RPE cells by staining PINK1 and PARKIN. One-year-old WT and dKO mice focusing on the RPE cells in the vicinity of the optic nerve (a,e). PINK1 (b, red) and PARKIN (c, green) were double-stained and the merged image (d) was used to count the colocalized puncta from WT. Similarly, in dKO PINK1 (f, red) and PARKIN (g, green) were double-stained, and the merged image (h) was used to count the colocalized puncta. dKO = NFE2L2/PGC1a double knockout. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32183173/) licensed under a CC-BY license.

Western blot image of PINK1 antibody (BC100-494) in multiple cells lines. Human HeLa (lane 1), Mouse NIH-3T3 (lane 2), L929 (lane 3) and Rat PC12 (lane 4) whole cell protein were separated by SDS-PAGE on a 7.5% polyacrylamide gel. Protein was transferred to PVDF membrane and probed with 2 ug/mL BC100-494 in 1% BSA and detected with an HRP-conjugated anti-rabbit secondary antibody using chemiluminescence. Observed molecular weight ~55 kDa (arrowhead).

Analysis of PINK1 in mouse liver and hypatocytes using PINK1 antibody. Observed molecular weight ~55 kDa. WB image submitted by a verified customer review.

Pathogenic mutants of Parkin are subjected to Ser65 phosphorylation. Phos-tag Western blotting for Parkin and Western blotting for PINK1 were performed using Parkin WT and a series of pathogenic mutants. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep01002) licensed under a CC-BY license.

Alteration of mitochondria and PD associated proteins in SH-SY5Y cells with telomere removal by CRISPR-Cas9. Representative Western blot of PGC-1alpha, NRF1, and PINK1 in SH-SY5Y cells transfected with either gTel or gCont (72 h). Beta-actin served as a loading control. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/18/10/2093), licensed under a CC-BY license.

35 ug Neuro2A whole cell lysate separated on 8% PAGE and stained for PINK1 (1:1000 in 5% milk powder in TBST; secondary antibody AP at 1:5000). WB image submitted by a verified customer review.

Rabbit heart tissue. IHC-P image submitted by a verified customer review.

HEK293 cells were co-transfected with PINK1 siRNA (#1 or #2) or scrambled siRNA (scrambled) and untagged wild-type (WT) or Ser65Ala (S65A) mutant Parkin as indicated using TransFectin reagent (Bio-Rad). 48 hrs post-transfection, cells were treated with/without 10 uM CCCP for 3 h. 0.25 mg of 1% Triton whole-cell lysate were subjected to immunoprecipitation with GST-Parkin antibody (S966C) covalently coupled to protein G Sepharose and then immunoblotted with anti-phospho-Ser65 antibody in the presence of dephosphorylated peptide. 5% of the IP was immunoblotted with total anti-Parkin antibody. 0.25 mg of whole-cell lysates were immunoprecipitated with anti-PINK1 antibody (S085D) and immunoblotted with anti-PINK1 antibody. Representative of three independent experiments. Image collected and cropped by CiteAb from the following publication (//rsob.royalsocietypublishing.org/cgi/doi/10.1098/rsob.120080) licensed under a CC-BY license.

Characterisation of mitochondrial content and function in PINK1-/- platelets. Cropped immunoblots showing PINK1 expression in 10 uM CCCP (6 hours) treated WT, but not KO platelets by IP (upper panel) with loading control of IP inputs by blotting for PINK1 (lower panel). Uncropped blots for PINK1 are provided in Supplementary Fig. S1b. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-32716-4), licensed under a CC-BY license.

Representative TEM picture. The red and green arrows indicate PINK and PARKIN immunogold particles in the dKO RPE samples, respectively (e). ML, melanosome. Scale = 0.2 um. Mitochondria (M). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32183173) licensed under a CC-BY license.

Protein content and oxidation. Gastrocnemius muscle was analyzed by immunoblotting (A) for oxidatively modified protein, STIM1, PARKIN, PINK1, PGC‐1 alpha , VDAC1, and 4HNE (B). Data are means ± SEM. *Significantly different from Tp53inp1+/+.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - PPM1H does not dephosphorylate Ser111 of Rab8A or key phosphorylation sites of AMPK & Akt signaling pathways.(A) HEK293 cells were transiently transfected with constructs expressing the indicated components. 24 hr post-transfection, cells were treated ±10 µM CCCP (Carbonyl cyanide m-chlorophenyl hydrazine) for 3 hr to induce activation of the PINK1 kinase & trigger Rab8A phosphorylation at Ser111 (Lai et al., 2015). (B) As in (A) except cells were immunoblotted with the indicated antibodies that recognize key phosphorylation sites of AMPK & Akt signaling pathways. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31663853), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - PINK1 & Parkin are phosphorylated after a decrease in delta Ψm in mouse primary neurons. Neurons were infected with lentivirus encoding PINK1-Flag (A), wild-type HA-Parkin (B) or HA-Parkin with either the S65A or S65E mutation (C). Cells were treated with the mitochondrial uncoupler CCCP (30 μm) for 1–3 h & subjected to SDS-PAGE in the absence or presence of 50 μm phos-tag. Note that mobility does not reflect the molecular weight of proteins in phos-tag PAGE (Kinoshita et al. 15), & thus, molecular weight markers are not shown in the bottom gels. The red & black asterisks in (C) indicate phosphorylation of Parkin at Ser65 & an additional minor phosphorylation site, respectively. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23751051), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - PINK1 translation is critical for the accumulation of PINK1 upon mitochondrial damage. aPINK1 mRNA levels were not altered by mitochondrial damage. HeLa cells were incubated with 10 μM CCCP or combination of 10 μM oligomycin & 4 μM antimycin for 2 h & then, PINK1 mRNA levels were measured by qRT-PCR. PINK1 mRNA level was normalized to the corresponding GAPDH levels & quantified as a percentage of control (n = 8, t-test). b, c Translation is involved in PINK1 accumulation upon CCCP treatment. HeLa cells were first pre-incubated with 100 μM cycloheximide (CHX) for 2 h & then washed. Cells were further incubated with 10 μM CCCP with or without CHX for 2 h (n = 4, two-way ANOVA). Values are mean ± SEM (n.s. = non-significant, ***p 1000 cells) & are shown as a percentage of control. Values are mean ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27456084), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - Inhibition of endogenous miR-27a/b function increases human PINK1 expression. a Schematic diagram of sequence alignment of miR-27a/b & anti-miR-27a/b. Asterisks (*) indicate conserved nucleotides. b Inhibition of endogenous miR-27a/b function increased luciferase activities in HeLa cells. Cells were transfected with the reporter constructs containing the full-length 3′UTR of human PINK1 mRNA with wild-type (WT) or mutated (Mut) seed match sites downstream of Renilla luciferase gene. 48 h post-transfection, luciferase activities were measured (n = 4, t-test). c-f Inhibition of endogenous miR-27a/b increased PINK1 protein levels in HeLa (C-D) & M17 cells (E-F). Cells were transfected with 150 nM of anti-miR-27a/b or anti-control (anti-Ctl). 48 h post-transfection, cells were harvested for Western blot. PINK1 protein level was normalized to corresponding GAPDH level & quantified as a percentage of control (n = 4, t-test). Data are shown as a percentage of control. Values are mean ± SEM (n.s. = non-significant, **p < 0.01, ***p < 0.00) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27456084), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - Regulation of PINK1 by miRNA. a Schematic diagram of AGO-specific RNA immunoprecipitation assay. b, cPINK1 mRNAs are associated with miRNA/RISC complex. HeLa cells were transfected with 50 nM of AGO2 siRNA or negative control. 48 h post-transfection, AGO2-bound mRNAs were pulled down from cell lysates with 2A8 anti-AGO2 antibody or non-specific IgG antibody. PINK1 mRNA levels were then measured by qRT-PCR. siRNA against AGO2 served as a quality control for AGO-specific RNA immunoprecipitation assay. PINK1 mRNA levels were quantified as a percentage of group 1 (n = 6, two-way ANOVA). d, e Knock-down of DICER1 increased PINK1 protein levels. HeLa cells were transfected with 50 nM DICER1 siRNA (DICER1 KD) or negative control (Ctl). 48 h post-transfection, cells were harvested for Western blot. The arrowhead indicates a full-length PINK1 band & the asterisk indicates a non-specific band. Each protein level was normalized to corresponding GAPDH level & quantified as a percentage of control (n = 4, t-test). Values are mean ± SEM (n.s. = non-significant, **p < 0.01, ***p < 0.001) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27456084), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - The expression levels of microtubule protein light chain 3 (LC3B), PTEN inducing putative kinase 1 (PINK1), Parkin, mitofusin 1 (Mfn1), toll-like receptor (TLR) 9, cytochrome c oxidase 4 (COX4), myeloid differentiation factor 88 (MyD88), & nuclear factor (NF)-kappa B in lung tissues from animals with spontaneous breathing (CON) or mechanical ventilation at high tidal volume (HTV) with saline, antimycin A (AmA) or cyclosporine A (CsA). (A) Levels of PINK1, Parkin, & Mfn1 mRNA. (B) Levels of LC3B, PINK1, Parkin, & Mfn1 protein by Western blot. (C) Relative expression of LC3B-II/LC3B-I & PINK1 protein. (D) Relative expression of Parkin & Mfn1 protein. (E) Levels of TLR9 & COX4 mRNA. (F) Levels of MyD88 & nuclear factor-kappa B (NF-kappa B) mRNA. (G) Levels of TLR9, COX4, MyD88, & NF-kappa B protein by Western blot. (H) Relative expression of TLR9 & COX4 protein. (I) Relative expression of MyD88 & NF-kappa B protein. Fold expression for target genes was normalized to that measured for the beta -actin gene. Both of these experiments were in triplicate. aP < 0.05 vs. CON group;bP < 0.05 vs. HTV group; & cP < 0.05 vs. AmA group. Image collected & cropped by CiteAb from the following publication (//www.frontiersin.org/article/10.3389/fimmu.2018.01477/full), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - Mitophagy contributes to removal of excess mitochondrial fragments in preeclampsia.(a, left panel) Representative TEM image of a cytotrophoblast from PTC & PE placenta collected at 29 weeks. Mitochondrial proximity to ER (MAM) is indicated by white arrows, mitochondrial fission events are depicted by white stars (scale bar: 500 nm; n = 8 separate PE placentae). (a, right panel) percentage of mitochondrial tethering to the ER in in PE vs. PTC (PE placentae, n = 8; PTC placentae, n = 7; unpaired Student’s t-test *P < 0.05). b Representative western blots for MFN2 & calreticulin in ER isolated from PE & PTC placentae (PE & PTC, n = 3 separate samples). c WB & associated densitometry of ASAH2 (normalized to TOM20) in mitochondria from PTC & PE placentae. d Representative TEM depicting mitophagy (white arrow) in cytotrophoblast cell from PE placenta (scale bar: 500 nm, n = 8 separate PE placentae). e Western blot & associated densitometry of PINK1 & Parkin in PE vs. PTC mitochondrial isolates. Densitometry for PINK1 blot was used to calculate the ratio of full-length PINK63kDa to cleaved PINK153kDa (PE & PTC, n = 4 separate placentae, *P < 0.05) Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41419-018-0360-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - Deficiency or mutant PINK1 reduces phosphorylation of LETM1 at Thr192. a, b Proteins extracted from PINK1 WT or KO MEFs (a) or mouse brain (b) were subjected to IP with anti-LETM1, probed with pT192 & reprobed with anti-LETM1 by WB. c Proteins extracted from human fibroblast of control (Con) or a PINK1-Q456X patient (Q456X) were subjected to IP with anti-LETM1, probed with pT192 & reprobed with anti-LETM1 by WB. d HEK293 cells were transfected with Adtrack GFP control, AdPINK1-WT, & AdPINK1-Q456X mutant for 1 day. Endogenous LETM1 protein was isolated by IP with anti-LETM1, probed with pT192 & reprobed with anti-LETM1 by WB. e SH-SY5Y cells were treated with 25 µM rotenone for 8 or 16 h. Total cell lysates were either analyzed by WB with anti-PINK1, or subjected to IP with anti-LETM1, probed with pT192 & reprobed with anti-LETM1 antibodies by WB. All above experiments were replicated three times, respectively Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29123128), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - Polyphyllin I suppressed, & PINK1 knockdown further suppressed, tumor growth in a MDA-MB-231 xenograft modelA. Tumor volumes were measured every week & differed at the end of the treatment period (*P < 0.01 compared to shCon cells treated with PPI, ##P< 0.01 compared to shPINK1 cells treated with PPI). B. Representative image of tumors from each group. C. Body weight changes in mice during the 6 weeks of PPI treatment. There were no differences in body weights between the PPI-treated shCon & vehicle-treated shCon groups or between the PPI-treated shPINK1 group & shCon groups. D. Representative tumor tissues were sectioned & subjected to H&E staining, TUNEL assay, & immunohistochemistry staining for C-CASP3. Scale bars: 50 μm. E. Representative tumor tissues from each group were prepared & subjected to western blot using anti-DRP1, -PARK2, -LC3B, -PINK1, & -C-CASP3 antibodies. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.14413), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - Corticosterone affects NIX-dependent mitophagy through decreasing PGC1 alpha in vivo.a–f Mice exposed to vehicle, corticosterone (10 mg/kg), corticosterone w/ phorbol 12-myristate 13-acetate (PMA pretreatment, 200 μg/kg), or PMA alone for 7 days. a Slide samples for IHC immunostained w/ LAMP1 (green), TOMM20 (red), & DAPI (blue). Scale bars, 100 μm (magnification, ×200). n = 5. b The expressions of NIX, PTEN-induced kinase 1 (PINK1), & BCL2 interacting protein 3 (BNIP3) detected w/ WB where beta -actin used as a loading control. n = 5. c Slide samples for IHC immunostained w/ synpatophysin (green), PSD95 (red), & DAPI (blue). Scale bars, 100 μm (magnification, ×200). n = 5. d Synaptophysin & PSD95 detected by WB. Loading control is beta -actin. n = 5. e The mice subjected to Y-maze test to evaluate spatial memory function. n = 6. f The mice subjected to forced swim test to evaluate depression-like behavior. n = 5. g Vehicle or RU 486 (5 mg/kg) injected mice presented w//w/out corticosterone (10 mg/kg) for 3 days. The expressions of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha ) & NIX visualized via WB. Loading control is beta -actin. n = 5. h The schematic model for mechanisms of inhibition in NIX-dependent mitophagy by glucocorticoid shown. All blots & IF images representative. n = 5 or 6 from each animal w/ two technical replicates each in results of IHC & WB. Quantitative data presented as a mean ± S.E.M. The representative images acquired by SRRF imaging system. Two-sided two-way ANOVA conducted except Fig. 8b, data of which analyzed by two-sided unpaired student’s t test. ** indicates p < 0.01 versus control & ## indicates p < 0.01 versus corticosterone, respectively. Data 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.

$Western Blot: PINK1 Antibody - BSA Free [BC100-494] - Polyphyllin I triggers PINK1/PARK2-dependent mitophagyA. MDA-MB-231 cells were treated with 8 μM PPI for different periods of time as indicated, & PARK2, P62, & ubiquitin (UB) levels in mitochondrial fractions were determined by western blot. B. Cells were cotransfected with GFP-UB & RFP-mito & treated with 8 μM PPI for 9 h, after which PARK2 (Alexa Fluor 405, pink) & P62 (Alexa Fluor 647, blue) immunostaining was detected using confocal microscopy. C. Cells cotransfected with GFP-UB & RFP-LC3 were treated with 8 μM PPI for 9 h, after which PARK2 (Alexa Fluor 405, pink) & TOMM20 (Alexa Fluor 647, blue) immunostaining was detected using confocal microscopy. Scale bars: 10 μm. D-E. MDA-MB-231 cells were treated with 8 μM PPI for different periods of time as indicated; whole-cell lysates were then separated on 8% SDS-PAGE gels & analyzed by western blot using the anti-PINK1 antibody. Relative full-length (∼63 kDa) & cleaved (∼52 kDa) PINK1 levels were quantified by densitometry & normalized to Tubulin. The results were expressed as a percentage of control, which was set at 100%. Data are presented as mean ± SD (*P< 0.01 vs. the control). F. Cells were treated with 8 μM PPI for different periods of time as indicated, & whole-cell lysates were then subjected to western blot analysis. G. Cells were treated with 8 μM PPI for 9 h, after which mitochondrial fractions were prepared & subjected to immunoprecipitation using anti-PINK1 antibody; associated PARK2 was detected using immunoblotting. H. RFP-mito-expressing MDA-MB-231 cells were treated with 8 μM PPI for 9 h, & PINK1 (Alexa Fluor 488, green) & PARK2 (Alexa Fluor 405, pink) immunostaining were evaluated using confocal microscopy. Scale bars: 10 μm. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.14413), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - miR-27a/b suppress human PINK1 expression. a Schematic diagram of the conserved target sites of miR-27a/b in the 3′UTR of human PINK1 mRNA. Seed sequences were indicated in red. Asterisks (*) indicate the conserved nucleotides. b Schematic diagram of the luciferase reporter plasmids. The reporter constructs contain the full-length 3′UTR of human PINK1 mRNA with wild-type (WT) or mutated (Mut) seed match sites downstream of Renilla luciferase gene. c Overexpression of miR-27a/b decreased luciferase activities in HeLa cells. Cells were transfected with miR-27a/b or negative control (Ctl) along with reporter constructs as indicated. 48 h post-transfection, luciferase activities were measured (n = 4, one-way ANOVA). Renilla luciferase activity was normalized to the corresponding firefly luciferase activity. d-g Overexpression of miR-27a/b decreased PINK1 protein levels in HeLa (D-E) & M17 cells (F-G). Cells were transfected with miR-27a/b or negative control (Ctl). 48 h post-transfection, cells were harvested for Western blot (n = 4, t-test). PINK1 protein level was normalized to corresponding GAPDH level & quantified as a percentage of control. Data are shown as a percentage of control. Values are mean ± SEM (n.s. = non-significant, **p < 0.01, ***p < 0.001) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27456084), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - Increased p62 expression is prevented in PINK1 knock‐down cells. (a) SH‐SY5Y cells expressing scrambled shRNA or PINK1 shRNA (117‐2, 118‐3, & 118‐4) were treated with 10 μM carbonyl cyanide m‐chlorophenylhydrazone (CCCP) for 24 h & full‐length (FL) PINK1 protein levels measured by western blot. No accumulation of FL PINK1 was observed in 118‐3 & 118‐4 PINK1 shRNA clones. (b) SH‐SY5Y cells constitutively expressing scrambled (scram) or PINK1 shRNA (clone 118‐3) were treated with vehicle or 10 μM CCCP for 24 h & p62 protein expression measured by western blotting. **p < 0.05 versus scram CCCP; n = 9. (c) SH‐SY5Y cells were incubated in serum‐free culture media for up to 24 h to induce starvation‐mediated macroautophagy. p62 protein levels were measured by western blotting. *p < 0.05 versus 0 h; **p < 0.01 versus 0 h; n = 4. (d) SH‐SY5Y cells were transfected with scrambled (scram) or p62 siRNA for 72 h. For the last 24 h, cells were treated with vehicle (ethanol) or 10 μM CCCP. p62 protein expression was then measured by western blotting. p62 protein was significantly decreased in p62 siRNA cells under basal conditions & following CCCP treatment, when compared to scram cells. **p < 0.01 versus scram vehicle or scram CCCP; n = 3. (e) Mitochondrial content was measured in SH‐SY5Y cells treated with scram or p62 siRNA for 72 h by measuring citrate synthase (CS) activity. For the last 24 h, cells were treated with vehicle or 10 μM CCCP & data are expressed as % of vehicle. *p < 0.05 versus scram CCCP treatment; n = 5. 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: PINK1 Antibody - BSA Free [BC100-494] - NF kappa B upregulates human PINK1 gene expression. (A) NF kappa B increases the endogenous human PINK1 mRNA level. HEK293 cells were transfected with either the NF kappa B expression vector or empty vector pMTF. RT-PCRs were performed using either primers specific to the human PINK1 coding sequence or the human beta -actin coding sequence. (B) NF kappa B increases the endogenous human PINK1 protein levels. HEK293 cells transfected with NF kappa B were harvested 48 h after transfection for protein detection. Cell lysates were run on 10% glycine gel & images were collected by Licor. A significant increase of endogenous PINK1 was observed. (C) The endogenous human PINK1 protein level was dramatically increased by NF kappa B in SH-SY5Y cells. SH-SY5Y cells were transfected with NF kappa B p65 plasmids or the control vector pMTF & then harvested for protein detections. The human PINK1 protein & its proteolysis products ∆1-PINK1 (55kD) & ∆2-PINK1 (45kD) were all increased. beta -actin acted as the internal control. (D) LPS treatment facilitated PINK1 expression in SH-SY5Y cells. Cells were harvested after being treated for 16 h & then subjected to Western blot. The human beta -actin level was served as a control. Quantification was performed by Image J software. Values indicate means ± SEM. n = 3, *p <0.01 by Student’s t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25108683), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - G-TPP induces PINK1 stabilization & kinase activity in HeLa cells(A) G-TPP treatment leads to PINK1 stabilization & pS65-Ub induction in HeLa cells. HeLa cells stably expressing untagged Parkin were treated with 10 µM G-TPP for the indicated times. Western blots were prepared with cell lysates & probed with antibodies against PINK1 & pS65-Ub. GAPDH served as a loading control. (B) pS65-Ub is induced in G-TPP treated cells & co-localizes with EGFP-Parkin & mitochondria. HeLa cells stably expressing EGFP-Parkin (green) were treated with 10 µM G-TPP for the indicated times & fixed. Cells were stained with antibodies against pS65-Ub (red) & the mitochondrial marker TOM20 (cyan). Scale bars correspond to 10 µM. (C) Quantification of Parkin translocation using High Content Imaging. HeLa EGFP-Parkin cells were treated for 4 or 8 h with or without 10 µM G-TPP. CCCP treatment (10 µM for 2 h) was used as a positive control. Cells were fixed, counterstained with Hoechst dye to visualize nuclei, imaged & analyzed using the ratio of cytoplasmic to nuclear EGFP signal [21]. Data was normalized to positive (2 h 10 µM CCCP treatment) & negative (2 h DMSO) controls. G-TPP significantly induced Parkin re-localization to levels similar to or beyond 2 h CCCP treatment. Shown are the mean values of three independent experiments with triplicate wells each ± SEM (one-way ANOVA with Tukey’s posthoc, ***p < 0.0005). Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.22287), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: PINK1 Antibody - BSA Free [BC100-494] - G-TPP leads to recruitment of autophagy adapters & degradation of mitochondria(A) HeLa cells stably expressing untagged Parkin were treated with 10 µM G-TPP for 8 h. Western blots were prepared from cell lysates & probed with antibodies against LC3, phospho-TBK1 (Ser172) & TBK1. GAPDH was used as a loading control. Upon 8 h the levels of LC3-I & LC3-II were both increased. At 8 h after treatment with G-TPP but not at 4 or 24 h, TBK1 was phosphorylated. (B) HeLa cells stably expressing EGFP-Parkin were treated with 10 µM G-TPP & fixed 8 h after treatment. Cells were stained with antibodies against the autophagy adapter proteins NBR1, NDP52, OPTN, p62, & TAX1BP1 (red). Mitochondria were counterstained with TOM20 antibodies (cyan), nuclei with Hoechst (blue). EGFP-Parkin epifluorescence is shown in green. Scale bar corresponds to 10 µM. (C) HeLa cells stably expressing EGFP-Parkin & the reporter protein mitoKeima were treated with 10 µM CCCP or G-TPP & imaged over time. The ratio of ‘neutral’ mitoKeima to ‘acidic’ mitoKeima was calculated as readout for mitophagy. Parkin translocation was monitored at the same time. Values for Parkin translocation & mitophagy were normalized to 12 h treatment with 10 µM CCCP as positive control & DMSO as negative control (two-way ANOVA with Tukey’s post-hoc test, **p < 0.005, ***p < 0.0005). Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.22287), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Rabbit Polyclonal PINK1 Antibody [BC100-494] - PINK1 detection in mouse and postmortem human cortex samples. 1:1000 dilution at 4 degrees overnight to detect endogenous PINK1 from mouse and postmortem human cortex samples (10 ug total protein). Image from a verified customer review.

Reactivity:

Hu, Mu, Rt, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, 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	BC100-494AF350
Alexa Fluor 405	BC100-494AF405
Alexa Fluor 488	BC100-494AF488
Alexa Fluor 532	BC100-494AF532
Alexa Fluor 594	BC100-494AF594
Alexa Fluor 647	BC100-494AF647
Alexa Fluor 700	BC100-494AF700
Alexa Fluor 750	BC100-494AF750
Allophycocyanin	BC100-494APC
Biotin	BC100-494B
CoraFluor 1	BC100-494CL1
DyLight 350	BC100-494UV
DyLight 405	BC100-494V
DyLight 488	BC100-494G
DyLight 550	BC100-494R
DyLight 594	BC100-494DL594
DyLight 650	BC100-494C
DyLight 680	BC100-494FR
DyLight 755	BC100-494IR
FITC	BC100-494F
HRP	BC100-494H
Janelia Fluor 525	BC100-494JF525
Janelia Fluor 549	BC100-494JF549
Janelia Fluor 646	BC100-494JF646
Janelia Fluor 669	BC100-494JF669
PE	BC100-494PE
PerCP	BC100-494PCP
mFluor Violet 450 SE	BC100-494MFV450
mFluor Violet 500 SE	BC100-494MFV500
mFluor Violet 610 SE	BC100-494MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	BC100-494SS

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NB100-417

Carbonic Anhydrase IX/CA9 Antibody - BSA Free

(7 Reviews)

(292 Publications)

0.1 ml

NB100-417

CA9 expressed in hypoxic regions as assessed by pimonidazole staining (A), but also observed in non-pimonidazole areas (B). Most of CA9 expressed in pimonidazole positive regions (C). Proliferation (BrdUrd) & apoptosis (caspase-3) in relation to CA9 expression shown in figure D & E. Red, CA9; Green, pimonidazole (A-B), BrdUrd (D) or caspase-3 (E); Yellow, overlap of CA9 (red) & pimonidazole (green); Light blue, vessels. Magnification 100x. Scale bars represent 100 um. Closed circles represent CA9 expression in pimonidazole positive regions; open circles represent CA9 expression in pimonidazole -ve regions. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0108068), licensed under a CC-BY license.

<b>Orthogonal Strategies Validation. </b>Formalin-fixed paraffin-embedded tissue sections of human stomach were probed for Carbonic Anhydrase IX/CA9 mRNA (ACD RNAScope Probe, catalog # 559348; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus catalog # NB100-417) at 1:1000 dilution with overnight incubation at 4 degrees Celsius followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.

Analysis on various human cell lysates. Specific bands were detected for Carbonic Anhydrase IX/CA9 in HEK-293 and A498 cell lines at a molecular weight of approximately 50 kDa. WB image submitted by a verified customer review.

Immunohistochemical representative microphotographs representing the HIF-1alpha, GLUT-1, and CAIX expression in endometrial cancer according to FIGO classification (IA, IB, II, IIIA, IIIC, and IV). Primary objective magnification 20x. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/bmri/2014/616850/) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Hypoxias impact on proteins of the mitochondrial ISC assembly machinery. (A) Immunoblotting analyzed the total protein extracts from HeLa cells grown in normoxia (Nx, 21% O2) or hypoxia (Hx, 1% O2) conditions using VDACs poly antibody and anti-CAIX, -ISCU, -FXN, -NFS1, -HSC20 antibodies. Beta-Actin was used as loading control. Citation: Ferecatu I, Canal F, Fabbri L, Mazure NM, Bouton C, Golinelli-Cohen M-P (2018) Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1 alpha activation. PLoS ONE 13(3): e0194782.

<b>Biological Strategies Validation. </b>Analysis using the DyLight 488 conjugate of NB100-417. Staining of Carbonic Anhydrase IX (red) in human glioma U87 cells. DAPI counterstains nuclei (blue). ICC/IF image submitted by a verified customer review.

Simple Western lane view shows a specific band for CAIX in 0.1 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

An intracellular stain was performed on U-87 MG Cells with NB100-417 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 APC-conjugated Secondary Antibody, (R&D Systems, F0111).

Western Blot analysis of Carbonic Anhydrase IX/CA9 Antibody on human breast cancer MCF7 cells. Image from verified customer review.

A431 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Carbonic Anhydrase IX/CA9 Antibody NB100-417 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.

An intracellular stain was performed on A431 cells with Carbonic Anhydrase IX/CA9 Antibody NB100-417 (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 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Analysis in 1) HeLa, 2) MDA-MB-231, and 3) A549 whole cell lysates. Specific bands were detected for Carbonic Anhydrase IX/CA9 at a molecular weight of 50 kDa.

Analysis on rat renal cortex. A specific band was detected at a molecular weight of approximately 50 kDa.

Immunocytochemical analysis of CNHCs with antibodies against the RCC marker CAIX. Clusters of CNHCs cytomorphologically classified as uncertain malignant (-UMF) with cytoplasmic positive staining with antibodies against the RCC marker CAIX. Image collected and cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-214) licensed under a CC-BY license.

Analysis of human colon carcinoma, xenografted in mice. IHC-Fr image submitted by a verified customer review.

IHC analysis of a FFPE tissue section of human breast cancer using CAIX antibody at 1:1000 dilution. The primary antibody bound to CAIX antigens in the tissue section was detected using a HRP labeled secondary antibody and DAB reagent. Nuclei of the cells were counterstained with hematoxylin. This CAIX antibody generated an expected cytoplasmic staining of CAIX protein with an intense signal around the cellular membranes in tumor cores. The latter are more likely to be hypoxic in growing tumors which signifies that the observed CAIX staining is specific.

Strong cytoplasmic and membranous CA9 staining in glandular cells of human stomach tissue. Citrate buffer pH 6 antigen retrieval. Antibody at 1:250 dilution and detected with an Alexa Fluor 546 labeled goat-anti-rabbit secondary antibody. Nuclei detected with DAPI. IHC-P image submitted by a verified customer review.

Immunofluorescence of human RCC tumor cryosections using NB100-417 (Panel A). Panel B shows staining with normal rabbit serum.

Colorectal cancer xenograft growth was suppressed by CB-PIC(20 and 50 mg/kg body weight) in female athymic nude mice. Starting three days after SW620 cell inoculation, CB-PIC (20 and 50 mg/kg body weight) was injected in abdomen with 4% Tween 20 as vehicle once daily. Representative examples of immunohistochemical staining for CA IX in tumor section. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/ecam/2013/974313/), licensed under a CC-BY license.

<b>Genetic Strategies Validation. </b>Iron depletion and nitric oxide stress and Iron depletion induce the truncated VDAC1 form accumulation. (D) Total protein extracts were analyzed by immunoblotting using VDACs poly antibody and anti-HIF-1 alpha and -CAIX antibodies. (E) Immunoblotting was used to analyze total protein extracts using antibodies against the three VDAC isoforms. (F) HeLa cells were grown in hypoxia (Hx, 1% O2) conditions and transfected with iscu- or NC-siRNA for 3 days, or grown in normoxia (Nx, 21% O2), some treated with DFO for 16 h. Western Blots analyzed total proteins using VDACs poly antibody and anti-HIF-1 alpha, -CAIX, -ISCU antibodies. Citation: Ferecatu I, Canal F, Fabbri L, Mazure NM, Bouton C, Golinelli-Cohen M-P (2018) Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1 alpha activation. PLoS ONE 13(3): e0194782.

The images are representative of immunohistochemistry staining of nuclear staining of proliferation marker Ki67, membrane protein CD 44, and CA IX. The images of scaffold culture (left panel) are compared with cell pellet paraffin section (right panel). The scaffold dissolved in xylene presented as a clear region marked ∗. All scale bars are 100 μm, magnification 40x.

Chemosensitivity of the CTC SCLC lines (IC50, mean ± SD) and immunohistochemistry of sections of tumorospheres. All differences between single cells (SC) and tumorospheres (TS) are statistically significant. Immunohistochemical staining of sections of UHGc5, BHGc16 and BHGc26 CTCs was performed using antibodies directed to CD56, CHGA, CAIX and Ki67, respectively.

Western Blot: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Analysis of protein levels of genes with increased transcription in hypoxia, IOX2 & VH032.HIF targets were increased in hypoxia, VH298 & FG-4592. 0.05% DMSO (vehicle control), 1% O2 (hypoxia), 100 µM VH298 & 50 µM FG-4592 were introduced to (A) HeLa or HFF for 24 hours & (B) HeLa for indicated time. Protein levels were analysed by immunoblotting using antibodies against indicated proteins, with beta -Actin as loading control. The blots shown are representative of three independent experiments. * indicates longer exposure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30801039), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - IGF1R & CA9 expression in PDAC.(A) Representative staining of CA9 cells quantified on a scoring system of 0–3 according to staining intensity (original magnification ×200). CA9 was mainly expressed in the cell membrane of PDAC cells. (B) IGF1R was mainly expressed in the cytoplasm of PDAC cells. (C) Association between IGF1Rexpression & CA9 expression in 120 pancreas cancer. A significant correlation was found between IGF1R expression level was significantly correlated with CA9 expression in PDAC cells (p < 0.01, rs = 0.649; Spearman’s rank sum test). The size of circle indicates the number of PDAC patients. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27487118), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Representative human colorectal cancer specimen. Immunohistochemistry of CA9 & GGT1 in mLNs. In these cases, both CA9 & GGT1 were expressed, suggesting that GGT1 expression is related to hypoxia in mLNs. Scale bar, 500 μm. Black arrowheads: GGT of cancer cells, white arrowheads: GGT surrounding cancer cells, arrow: accumulation of GGT. mLNs: metastatic lymph nodes, GGT: gamma-glutamyl transpeptidase. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30542087), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Representative microscopy images of staining for hypoxia markers in prostate tissues (MO, 400×). A) HIF-1 alpha - notice the granular cytoplasmic immunoreactivity of the malignant epithelial cells. In this case, more than 50% of the glands stained. B) LOX - strong & diffuse nuclear immunoreactivity of the epithelial cells. C) CAIX - note a focal apical cytoplasmic immunoreactivity in epithelial cells. D) VEGFR2 - moderate nuclear & weak cytoplasmic expression of the epithelial cells Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28143503), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - The images are representative of immunohistochemistry staining of nuclear staining of proliferation marker Ki67, membrane protein CD 44, & CA IX. The images of scaffold culture (left panel) are compared with cell pellet paraffin section (right panel). The scaffold dissolved in xylene presented as a clear region marked ∗. All scale bars are 100 μm, magnification 40x. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24101930), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Hypoxia, blood vessel & proliferation staining. IHC staining in the Biopsy 23 & Biopsy 3 respectively for CA-9, EF5, CD31, Ki67 & LYVE1. 15× Magnification. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/2072-6694/4/3/821), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - VH298 elicits a HIF-dependent hypoxic response in various cell lines.1% DMSO, hypoxia (1% O2) & indicated compounds at 100 μM were introduced to (a) HFF, HeLa, U2OS & (b) RCC4 cells expressing HA-tagged VHL (with the exception of cis-VH298 & VH298 used at 150 μM) for 24 h & (c) CTLs for 8 h. Protein levels were analysed by immunoblotting using antibodies against the indicated proteins, with beta -actin (SMC1 for CTLs) as the loading control. The blots shown are representative of three independent experiments. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27811928), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - HE staining & immunohistochemistry (CA9 & GGT1) of fixed primary tumor & mLN specimens from orthotopic mouse model. CA9 & GGT1 were expressed inside the primary tumor & in mLNs. GGT1 was accumulated in the area of central necrosis. HE: hematoxylin-eosin. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30542087), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Comparison between primary tumor & mLNs of the mouse model. Expression of CA9 & GGT1 in mLNs was higher than that in the primary tumor at 1week after injection of cells. (A) CA9 & GGT1 expression of HT29 mLNs compared to primary tumor at 6 weeks or 1 week after injection of cells. (B) Tissue oxygen concentration (%) of rectal submucosa (SM) & nLNs (n = 3). Error bars represent SD. mNs: metastatic lymph nodes, nLNs: non metastatic lymph nodes. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30542087), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Chemosensitivity of the CTC SCLC lines (IC50, mean ± SD) & immunohistochemistry of sections of tumorospheres. All differences between single cells (SC) & tumorospheres (TS) are statistically significant. Immunohistochemical staining of sections of UHGc5, BHGc16 & BHGc26 CTCs was performed using antibodies directed to CD56, CHGA, CAIX & Ki67, respectively. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-05562-z), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Immunocytochemical analysis of CNHCs with antibodies against the RCC marker CAIX. Clusters of CNHCs cytomorphologically classified as uncertain malignant (−UMF) with cytoplasmic positive staining with antibodies against the RCC marker CAIX (A). Clusters of CNHC-UMF & -BF without reactivity for CAIX antibodies (B & C, respectively). A single CNHC-MF with positive cytoplasmic (D) & without staining for CAIX (E). Single CAIX-negative CNHC-UMF & -BF (F & G, respectively). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-214), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - GPRC5A promotes hypoxic cell survival via YAPAHypoxia induced increased total YAP expression in a panel of colorectal cancer cell lines. Actin confirmed equal loading; blots are representative of at least two independent experiments. Related to Fig 4.B–EqRT–PCR analysis revealed that hypoxia increased the expression of known YAP target genes AREG, BCL2L1 (BCL‐XL), CTGF & CYR61. Representative experiments are shown. Representative examples of n = 3 independent experiments are shown; data are presented as mean ± SD.FIncreased expression of YAP Ser397 in GPRC5A‐depleted cells was overridden by expression of constitutively active RhoA (G14V) in hypoxia. Expression of dominant negative RhoA (T19N) did not further increase YAP Ser397 phosphorylation in response to GPRC5A depletion.GDarker exposure of the GPRC5A blot in Fig 4I, confirming expression of 30–40 kDa & 80 kDa species. Note that YAP siRNA partially diminishes GPRC5A in line with the existence of positive HIF‐GPRC5A‐YAP feedback loop. Asterisk (*) indicates non‐specific band.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30143543), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - CAIX expression in PDAC & association with outcomeA. Representative images of immunohistochemical staining showing: negative expression of CAIX in both tumor cell & stromal compartments (top left), positive expression of CAIX in tumor cells but not stromal compartment (top right), positive expression of CAIX in stromal compartment but not tumor cells (bottom left), positive expression of CAIX in both tumor cell & stromal compartments (bottom right). B. Kaplan-Meier analysis of CAIX expression in the distinct tumor cell & stromal compartments & the associated overall survival. C. Representative images of immunohistochemical staining showing low stromal volume & high stromal volume & low & high microvessel density D. Representative images of immunohistochemical staining showing: positive expression of MCT4 in tumor cell compartment & positive expression of MCT4 in tumor stromal compartment. Kaplan-Meier analysis of combinatorial MCT4 & CAIX expression in the stromal compartment & the associated overall survival. Hazard ratios & p-values for each comparison are summarized in the table. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27623078), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Hypoxia & autophagy markers in breast cancer stem-cells(A) Co-expression of CD133 & BECLIN1 in pCR & non-pCR patients. Double IF X800. (B) Tumor cells (T) distributed around necrosis (N) express LC3B with typical cytoplasmic punctuated staining, ×1000. Double IF shows that some CD133-positive tumor cells co-express LC3B. ×400. (C) Tumor cells (T) distributed around necrosis (N) express CAIX, on their cytoplasmic membrane, ×1000. Double IF shows a co-expression of LC3B & CAIX in some tumor cells. ×400. (D) Laser microdissection of CD133-positive cells. (E) Laser-microdissected CD133-positive cells express PROMININ & not PTPRC (CD45). (F) mRNA relative quantification of HIF1A, CAIX, LC3B, BECN1, & BNIP3L show a significantly higher expression level in CD133-positive cells versus CD133-negative cells, in the 35 non-pCR biopsies. *p < 0.05. (G) Multivariate analysis of tumor cell characteristics & pCR shows an inverse correlation between pCR & cells bearing markers of stemness or autophagy. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28445132), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Immunofluorescent images showing CAIX expression, pimonidazole, apoptosis & proliferation in SCCNij202.CAIX is expressed in hypoxic regions as assessed by pimonidazole staining (A), but is also observed in non-pimonidazole areas (B). Yet, most of CAIX is expressed in pimonidazole positive regions (C). Proliferation (BrdUrd) & apoptosis (caspase-3) in relation to CAIX expression are shown in figure D & E respectively. Red, CAIX; Green, pimonidazole (A–B), BrdUrd (D) or caspase-3 (E); Yellow, overlap of CAIX (red) & pimonidazole (green); Light blue, vessels. Magnification 100×. Scale bars represent 100 µm. Closed circles represent CAIX expression in pimonidazole positive regions; open circles represent CAIX expression in pimonidazole negative regions. Abbrevations: 1× S4, 8 h, one i.p. injection S4, harvest after 8 hours; 1× S4, 24 h, one i.p. injection S4, harvest after 24 hours; 3× S4, one i.p. injection S4 a day for 3 days, harvest 8 hours after the last injection; 5× S4, one i.p. injection S4 a day for 5 days, harvest 8 hours after the last injection. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25225880), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Colorectal cancer xenograft growth was suppressed by CB-PIC(20 & 50 mg/kg body weight) in female athymic nude mice. Starting three days after SW620 cell inoculation, CB-PIC (20 & 50 mg/kg body weight) was injected in abdomen with 4% Tween 20 as vehicle once daily. (a) Body weights of mice. (b) Tumor growth in a time course. (c) Final tumor weight at termination of experiment. (d) Representative examples of immunohistochemical staining for Ki-67, CD34, TUNEL, CA IX & pAMPK alpha in tumor sections. Graphs show the Ki67 index (proliferation), CD34 index (angiogenesis), TUNEL index (apoptosis), CA IX (hypoxia region), & pAMPK alpha index in tumor sections. Values are means ± SD, n = 6. *P < 0.05 & ***P < 0.001 compared with control mice. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23589723), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Hypoxia induces GPRC5A directly via HIFsASILAC‐based proteomics data identify known (red) & novel (green) hypoxia‐induced proteins in SW620 cells. One‐sample t‐test was performed.BWestern blotting confirmed GPRC5A as a hypoxia‐induced protein in SILAC lysates.CValidation of GPRC5A Western blot data using siRNA. *Non‐specific band of ˜60 kDa not depleted by GPRC5A siRNA.DConfocal microscopy showing plasma membrane GPRC5A expression in hypoxic SW620 cells (scale bars: 75 μm).EWestern blotting showing GPRC5A upregulation by hypoxia in a panel of colorectal tumour cell lines.FBasal & hypoxia‐induced GPRC5A protein expression was decreased by HIF‐1/2 alpha depletion.GDepletion of HIF‐1 beta decreased GPRC5A protein upregulation in hypoxia.HHypoxia mimetic DMOG induced HIF‐1/2 alpha , CA9 & GPRC5A protein expression. Dual HIF‐1/2 alpha depletion reduced GPRC5A induction by DMOG.IqRT–PCR demonstrating that GPRC5A mRNA was upregulated by hypoxia (n = 3). GPRC5A was normalised to HPRT (error bars ± SD).JqRT–PCR demonstrating that HIF‐1/2 alpha depletion decreased GPRC5A induction during hypoxia (n = 3). GPRC5A was normalised to HPRT (error bars ± SD).KChIP‐PCR analyses identify HIF‐1 alpha binding to the GPRC5A promoter region containing a putative optimal HRE (error bars ± SD, n = 3).Data information: Asterisks (*) indicate non‐specific band. Level adjustments were made to images in Adobe Photoshop post‐acquisition for clarity (equal changes applied to the entire image). Representative examples of n = 3 independent experiments are shown.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30143543), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Carbonic Anhydrase IX/CA9 Antibody - BSA Free [NB100-417] - Establishment of a human protein replacement stable cell line using 47snoMEN.(a) Images of protein replacement stable cell lines (U2OSGFP–HIF-1 alpha -47PR). Expression of FP proteins was confirmed by fluorescence imaging with DFX treatment. Bar is 10 μm. (b) U2OS & U2OSGFP–HIF-1 alpha -47PR cells were exposed to 1% O2 for the indicated periods of time prior to lysis. Cell extracts were analysed by western blot using the indicated antibodies. (c) U2OS cells were transfected with 1 μg of control & HIF-1 alpha -47snoMEN vectors for 48 hours prior to lysis. DFX was added for the last 24 hours. Cell extracts were analysed by western blot using the indicated antibodies. (d) 3D culture analysis. HeLaGFP–HIF-1 alpha -47PR cells were cultured in lipidure coated plates to induce spheroid formation. HIF-1 alpha is visible in the core of the spheroid revealing hypoxia. Bar is 10 μm. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0154759), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Pl

Species Glossary

Reactivity: Hu, Mu, Rt, Pl

Applications:

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

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

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Rabbit Polyclonal

Host: Rabbit Polyclonal

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Sample Size Available

Sizes: Sample Size Available

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Unconjugated

Available Conjugates:

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Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-417AF350
Alexa Fluor 405	NB100-417AF405
Alexa Fluor 488	NB100-417AF488
Alexa Fluor 532	NB100-417AF532
Alexa Fluor 594	NB100-417AF594
Alexa Fluor 647	NB100-417AF647
Alexa Fluor 700	NB100-417AF700
Alexa Fluor 750	NB100-417AF750
Allophycocyanin	NB100-417APC
Allophycocyanin/Cy7	NB100-417APCCY7
Biotin	NB100-417B
CoraFluor 1	NB100-417CL1
DyLight 350	NB100-417UV
DyLight 405	NB100-417V
DyLight 488	NB100-417G
DyLight 550	NB100-417R
DyLight 594	NB100-417DL594
DyLight 650	NB100-417C
DyLight 680	NB100-417FR
DyLight 755	NB100-417IR
FITC	NB100-417F
HRP	NB100-417H
Janelia Fluor 525	NB100-417JF525
Janelia Fluor 549	NB100-417JF549
Janelia Fluor 646	NB100-417JF646
Janelia Fluor 669	NB100-417JF669
PE	NB100-417PE
PE/Atto594	NB100-417PEATT594
PE/Cy5.5	NB100-417PECY55
PE/Cy7	NB100-417PECY7
PerCP	NB100-417PCP
mFluor Violet 450 SE	NB100-417MFV450
mFluor Violet 500 SE	NB100-417MFV500
mFluor Violet 610 SE	NB100-417MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-417SS

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DY417

Mouse IL-10 DuoSet ELISA, 15 Plate

(307 Publications)

1 Kit (for 5 Plates)

DY417

Reactivity:

Mu

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Reactivity: Mu

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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

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