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NB100-105

HIF-1 alpha Antibody (H1alpha67)

(45 Reviews)

(1013 Publications)

0.1 ml

NB100-105

HIF-1 alpha induction by CoCl2 on Caki-1 cell lysate. WB image submitted by a verified customer review.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>HIF-1 alpha was detected in immersion fixed DFO treated Hela cells (left) but was not detected in HIF-1 knockout HeLa cells (right) using Mouse Anti-human HIF-1 alpha monoclonal antibody (Catalog # NB100-105) at 25 ug/mL for 3 hours at room temperature. Cells were stained using a NorthernLights (TM) 557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Naive CD4 T cells from WT, VHL-deficient (Vhl KO), or HIF-1 alpha-deficient (HIF-1 alpha KO) mice were differentiated under IL-22-skewing conditions for a total of 60 h. Some cells remained at normoxia for the duration of the culture (N); others were at normoxia for 35 h and then hypoxia (1% O2) for 24 h (H). At 60 h, nuclear extracts were harvested, and HIF-1 alpha and Lamin B1 levels were analyzed by Western blot. Image from verified customer review.

Analysis using the Alexa Fluor (R) 488 conjugate of NB100-105. Staining of HIF-1 alpha in multiple myeloma cells: H929 cells (0.5 x 10^6) were stained with Alexa Fluor 488(R) conjugated HIF-1 alpha antibody (NB100-105AF488). Image courtesy of Dr. Barbara Muz at Washington University in St. Louis School of Medicine.

Staining of HIF-1 alpha in human kidney using NB100-105. Renal tubular epithelium showed moderate membranous, cytoplasmic and nuclear staining, and glomeruli showed faint to moderate nuclear staining.

HIF1alpha/PFKFB3 is upregulated in beta-cells of HIP rats and humans with type 2 diabetes. Representative Western blot of PFKFB3 and HIF1alpha levels in nuclear-enriched- and whole cell extracts from non-diabetic (ND) and T2D donor islets. Data are presented as mean +/- SEM, n = 3 independent biological samples for each group. Statistical significance was analyzed by Student t-test (*p 80% of endogenous Sin1 while levels of myc-FL Sin1 associated with Rictor are comparable with endogenous Sin1 levels. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.20086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

GRPs enhance axonal growth in DRGs in vitro. A–C, Representative montage images are shown for control dissociated DRG culture (A), dissociated DRGs cocultured with rat GRPs (B), and dissociated DRGs cultures exposed to conditioned medium from alkaline phosphatase-expressing rat GRPs (C). beta III-tubulin (red) immunofluorescence highlights the neurons; GRPs are visualized by immunostaining for alkaline phosphatase (green). D shows quantification for the average length of the longest axon per neuron ± SEM (n ≥ 30 neurons in three separate experiments; **p ≤ 0.01 by Student’s t test). Scale bar, 250 μm. Image collected and cropped by CiteAb from the following open publication (//www.eneuro.org/lookup/doi/10.1523/ENEURO.0171-16.2017), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Vps13 co-fractionates with Rab7 and Rab5.(A) Western blot analysis of control fly head samples fractionated into a cytosolic and membrane fraction from postnuclear supernatant (PNS). EGFR was used as a membrane marker and GAPDH as a cytosolic marker. (B) Membrane fractions from control fly heads treated with 1 M KCl, Na2CO3 pH 11 or 6 M urea were centrifuged to separate the soluble and insoluble (membrane containing) fractions. The level of Vps13 was determined in these fractions. Markers for peripheral membrane proteins (GM130), integral membrane proteins (EGFR) and the cytosolic proteins (GAPDH) were used. The “Vps13 lysate” lane contains a lysate derived from Vps13 homozygous mutant fly heads, as expected no Vps13 is detected, demonstrating the specificity of the antibody against Vps13. (C) Membranes from control fly heads were fractionated on a sucrose gradient. Western blot analysis was performed to analyze the distribution of Vps13 in relation to markers associated with membranes of various organelles: Rab7 (late endosomes), Rab5 (early endosomes), GM130 (golgi), Lamp1 (lysosomes) and ATP5A (mitochondria). (D) Immunoisolation of membranes from fraction 14 of the sucrose gradient using Vps13 NT, Rab7 and Rab5 antibodies. (E) Quantification of the sucrose gradient fractionation of Fig 2C. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28107480), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Inhibition of CIP2A by siRNA impeded cell viability and DNA synthesis in HPV‐16E6–expressing cells. A, Elevated expression of CIP2A protein in 16E6‐expressing RPE1 cells. B, Western blot analysis of CIP2A and p53 proteins after transfection with scrambled siRNA (siCon) or CIP2A siRNA (siCIP2A) for 48 h. A representative of 3 independent experiments is shown. C, Cell viability assay of RPE1‐16E6 cells with CIP2A knockdown. D, Representative flow cytometry of BrdU staining profiles is shown. E, The mean and SD of BrdU‐positive cells from 3 experiments are summarized. **, P 30°. Scale bars represent 5 μm.(B) Representative graph of delta KT, delta MT, and the difference between delta KT and delta MT through time in a lateral kinetochore assessed from movie stills as in (A).(C) Graph of attachment state transitions experienced by percentage of lateral kinetochores, within a 50 s window, measured from time-lapse movies of cells transfected with siRNA oligos as indicated and treated with monastrol for 1 hr prior to imaging.(D) Graph of interkinetochore distances describing fate of three randomly chosen kinetochores from conditions described in (A).(E) Cumulative graph and T50 table showing lifetime of lateral kinetochores in HeLa cells treated with either control or MCAK siRNA or HeLa cells expressing tetracycline-inducible siRNA-resistant HEC1Ndc80-delta loop-YFP as indicated. The dashed line marks T50, the period when 50% of lateral kinetochores had ceased to remain laterally attached.(F) Representative immunofluorescence images of control or MCAK siRNA-transfected cells treated with monastrol for 1 hr prior to fixation and immunostained with antibodies against astrin and tubulin and CREST antisera and stained with DAPI for DNA. Insets are 3× magnified. Scale bars represent 5 μm (insets, 2 μm).(G) Graph of the percentage of lateral kinetochore pairs with both, one, or none of the pair displaying astrin in cells treated as in (F).(H) 3D-rendered images are rotated (as indicated) showing a lateral kinetochore in MCAK siRNA-treated cells immunostained with antibodies against astrin (red) and beta -tubulin (green) and CREST antisera (blue). The blue arrow marks kinetochore attachment site and pink arrow marks the plus end of laterally interacting microtubules. Scale bars represent 2 μm.p values representing significance levels were obtained with the Mann-Whitney U test (C), proportion test (D and F), and paired sample t test (E). ∗ and # indicate significant and insignificant differences, respectively. See also Figure S4. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23891108), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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) and 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) and 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 and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.ppat.1006262), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ca, Fe, Ma, Pm, Pm, Rb, Sh, Xp

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Fe, Ma, Pm, Pm, Rb, Sh, Xp

Applications:

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

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

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-105AF350
Alexa Fluor 405	NB100-105AF405
Alexa Fluor 488	NB100-105AF488
Alexa Fluor 532	NB100-105AF532
Alexa Fluor 594	NB100-105AF594
Alexa Fluor 647	NB100-105AF647
Alexa Fluor 700	NB100-105AF700
Alexa Fluor 750	NB100-105AF750
Allophycocyanin	NB100-105APC
Biotin	NB100-105B
DyLight 350	NB100-105UV
DyLight 405	NB100-105V
DyLight 488	NB100-105G
DyLight 550	NB100-105R
DyLight 594	NB100-105DL594
DyLight 650	NB100-105C
DyLight 680	NB100-105FR
DyLight 755	NB100-105IR
FITC	NB100-105F
HRP	NB100-105H
Janelia Fluor 549	NB100-105JF549
Janelia Fluor 646	NB100-105JF646
PE	NB100-105PE
PerCP	NB100-105PCP
mFluor Violet 450 SE	NB100-105MFV450
mFluor Violet 500 SE	NB100-105MFV500
mFluor Violet 610 SE	NB100-105MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Azide and BSA Free	NBP2-80760

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NBP2-46892

LC3B Antibody (1251A) - BSA Free

(3 Reviews)

(12 Publications)

0.1 mg

NBP2-46892

Western Blot: LC3B Antibody (1251A) [NBP2-46892] - The expression of LC3B in rat tissue. This image was submitted via customer Review.

<b>Biological Strategies Validation. </b>Flow (Intracellular): LC3B Antibody (1251A) [NBP2-46892] - Jurkat cells were either untreated (A) or treated with 50uM chloroquine for 24 hours (B). An intracellular stain was performed with anti-LC3B (1251A) antibody [Catalog # NBP2-46892] (blue) and a matched isotype control [Catalog # MAB1050] (orange). Cells were fixed with 4% paraformaldehyde, following fixation, cells were permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by rabbit IgG APC-conjugated secondary antibody (F0111, R&D Systems).

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Effects of PDTC on LC3 on the hippocampus of rats harvested 12 h after CLP. Values of protein data are normalized to actin level and given in relative arbitrary units. LC3-II increased in CLP rats receiving vehicle or PDTC compared with those in sham-operated rats. PDTC + CLP rats showed an increase in LC3-II compared with vehicle-treated rats. Data were expressed as mean +/- SEM, n = 6/group. *P < 0.05 vs sham-operated group. # P < 0.05 vs 12 h or Veh + CLP group. PDTC pyrrolidine dithiocarbamate, LC-3 microtubule-associated protein light chain-3, CLP cecal ligation and puncture. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26067996) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: LC3B Antibody (1251A) [NBP2-46892] - HeLa cells were treated with Chloroquine for 24 hours prior to fixation, permeabilization and incubation with anti- [Catalog # NBP2-46892] and anti tubulin (NB100-690) antibodies. Image enlargement shows the accumulation of LC3 (green) on autophagosomes in response to Chloroquine treatment. Tubulin staining is shown in red and DNA is counterstained with DAPI (blue).

Immunohistochemistry-Paraffin: LC3B Antibody (1251A) [NBP2-46892] - IHC (Immunohistochemical) analysis of a formalin fixed and paraffin embedded tissue section of normal mouse brain using rabbit monoclonal LC3B (1251A) antibody [Catalog # NBP2-46892] at 1:100 dilution with HRP-DAB detection. The antibody generated a weak diffused cytoplasmic staining in most of the cells but some cells, especially within empty areas on the section, showed punctate signal also which signifies the presence of autophagy in those areas.

Flow Cytometry: LC3B Antibody (1251A) [NBP2-46892] - HeLa cells untreated or treated with 50 uM chloroquine for 24h. Image from the Alexa Fluor 405 version of this antibody.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Analysis shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with Chloroquine. PVDF membrane was probed with 0.2 ug/mL rabbit anti-LC3B monoclonal Antibody (1251A) (Catalog # NBP2-46892, Novus Biologicals), followed by 1:2000 dilution of goat anti-rabbit IgG secondary antibody. A specific band was detected for LC3B at a molecular weight of approximately 15 kDa in CQ treated NIH-3T3 and PC12 cell lines.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Western Blot analysis using HeLa cells treated with (+) or without (-) Chloriquine (CQ). A specific band was detected for LC3B at a molecular weight of approximately 15 kDa.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Western Blot image of monoclonal anti-LC3B Antibody (Clone 1251D) [Catalog # NBP2-46892]. HeLa and Neuro2A cells were treated with or without 50 uM chloroquine for 24 hours as indicated. Whole cell protein was then separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane for 30 min and blocked in 5% non-fat milk in TBST (Tris-buffered saline, 0.1% Tween 20). The membrane was probed with 2 ug/ml anti-LC3B Antibody in 1% milk, and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the accumulation of LC3B II upon chloroquine treatment. Bands for LC3 were detected at a molecular weight of approximately 15 kDa in treated HeLA cells, and both treated and untreated Neuro2A cells.

Immunocytochemistry/Immunofluorescence: LC3B Antibody (1251A) [NBP2-46892] - HeLa cells were treated with 50 uM Chloroquine for 24 hour prior to fixation in 10% buffered formalin for 10 min. Cells were permeabilized in 0.1% Triton X-100 and incubated with 20 ug/ml anti- [Catalog # NBP2-46892] and 1:500 anti-tubulin [Catalog # NB100-690] for 1 h at room temperature. LC3B reactivity (green) was detected with ant-rabbit Dylight 488 and tubulin (red) with anti-mouse Dylight 550. Nuclei were counterstained with DAPI (blue).

Flow (Intracellular): LC3B Antibody (1251A) [NBP2-46892] - HeLa human cervical epithelial carcimoa cell line either treated with 50 uM chloroquine for 24 hours (filled histogram) or untreated (open histogram) was stained with Rabbit anti-Human LC3B Alexa Fluor 647 conjugated monoclonal antiobdy (1251A) [Catalog # NBP2-46892AF647]. To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation and Permeabilization Buffer Kit (FC012-NOV).

Immunoprecipitation: LC3B Antibody (1251A) [NBP2-46892] - Western blot analysis of LC3 immunoprecipitation. 10 ug of monoclonal anti-LC3B antibody (clone 1251A) [Catalog # NBP2-46892] was used to immunoprecipitate LC3 from 200 ug of total protein from HeLa cells treated with or without 50 uM chloroquine for 24 h. Antibody-protein was captured by magnetic protein A/G beads, washed and subjected to SDS-PAGE on a 4-15% gel. Protein was transferred to 0.2 um PVDF membrane and probed with 2 ug/ml anti-LC3B (NB100-2220) and detected with an anti-rabbit HRP conjugated secondary antibody using chemiluminescence. The detection of LC3 at a molecular weight of approximately 12 kDa and the antibody heavy chain (Ab Hc) at a molecular weight of approximately 50 kDa is indicated.

Reactivity:

Hu, Mu, Rt, Po, Bt, Bv, Fi, Xp, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bt, Bv, Fi, Xp, Ze

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-46892AF350
Alexa Fluor 405	NBP2-46892AF405
Alexa Fluor 488	NBP2-46892AF488
Alexa Fluor 532	NBP2-46892AF532
Alexa Fluor 594	NBP2-46892AF594
Alexa Fluor 647	NBP2-46892AF647
Alexa Fluor 700	NBP2-46892AF700
Alexa Fluor 750	NBP2-46892AF750
Allophycocyanin	NBP2-46892APC
Biotin	NBP2-46892B
DyLight 350	NBP2-46892UV
DyLight 405	NBP2-46892V
DyLight 488	NBP2-46892G
DyLight 550	NBP2-46892R
DyLight 594	NBP2-46892DL594
DyLight 650	NBP2-46892C
DyLight 680	NBP2-46892FR
DyLight 755	NBP2-46892IR
FITC	NBP2-46892F
HRP	NBP2-46892H
Janelia Fluor 549	NBP2-46892JF549
Janelia Fluor 646	NBP2-46892JF646
PE	NBP2-46892PE
PerCP	NBP2-46892PCP
mFluor Violet 450 SE	NBP2-46892MFV450
mFluor Violet 500 SE	NBP2-46892MFV500
mFluor Violet 610 SE	NBP2-46892MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-46892SS
Azide and BSA Free	NBP2-80829

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NB110-53818

ATG5 Antibody - BSA Free

(7 Reviews)

(266 Publications)

0.1 ml

NB110-53818

CQ increased CDDP-induced cell death in drug-resistant EOC through autophagy inhibition and induction of p21WAF1/CIP1.a A2780-CP20 cells stably expressing control or ATG5 shRNA were treated with CDDP for 48 h, and the expression of ATG5 and autophagy marker proteins, LC3 and p62, were analyzed by western blot using specific antibodies. Knockdown efficiency of ATG5 was shown by western blot. Expression of LC3 and p62 was also analyzed by western blot in A2780-CP20 cells treated with CDDP, CQ, or the CQ-CDDP combination for 48 h. beta-actin was used for a protein-loading control. shRNA/CP20 represents control-shRNA- or ATG5-shRNA-expressing A2780-CP20 cells. Western blot was performed three times, and representative data are presented (left panel). The amounts of p62 and LC3-I and -II normalized to the amount of beta-actin are represented by a bar graph (middle and right panels, respectively). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33277461), licensed under a CC-BY licence.

FIGURE 2: Blockage of autophagy leads to increased plp mRNA and protein levels. (B) Western blot analysis with antibodies against Atg5, P62, PLP and MBP proteins in optic nerve lysates of 6mo control and cKO mice. Representative images of a single experiment are depicted. Image collected and cropped by CiteAb from the following publication (//www.cell-stress.com/researcharticles/2022a-ktena-cell-stress/), licensed under a CC-BY licence.

LHPP suppresses aerobic glycolysis.A Relative mRNA expression levels of glycolytic genes and glutamine transporters in LHPP-overexpressing or LHPP-knockdown MGC-803 cells and control cells by qPCR. B Protein expression levels of glycolytic genes and glutamine transporters in LHPP-overexpressing or LHPP-knockdown MGC-803 cells and control cells on western blot. C IHC staining of LHPP and HIF1A in TMAs and their correlation. Scale bars = 100 μm. D Oxygen consumption rate and extracellular acidification rate of LHPP-overexpressing or LHPP-knockdown MGC-803 cells and control cells were measured using the Seahorse Bioscience XF96 analyser. E Human phosphokinase microarray assay analysis of the conditioned medium from stably transfected HGC-27 cells. A summary of the relative signal intensities of the indicated proteins is shown. G, H IHC staining of LHPP and p-GSK3b in TMAs and their correlation. Scale bars = 200 μm. F Combined LHPP and GSK3b by co-immunoprecipitation. I Protein expression levels of the Wnt pathway in LHPP-overexpressing or LHPP-knockdown MGC-803 cells and control cells on western blot. Data were analyzed using the Wilcoxon test. *P < 0.05, **P < 0.01, ***P < 0.001, KEGG Kyoto encyclopaedia of genes and genomes, GO gene ontology, qPCR quantitative polymerase chain reaction, IHC immunohistochemical, TMA tissue microarray. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35568711), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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

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 and diffuse nuclear immunoreactivity of the epithelial cells. C) CAIX - note a focal apical cytoplasmic immunoreactivity in epithelial cells. D) VEGFR2 - moderate nuclear and weak cytoplasmic expression of the epithelial cells Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28143503), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Interaction of Nesprin-2-SMC and Nesprin-2 with SMC2 and SMC4. (a) Precipitation of SMC2 and SMC4 with GST-Nesprin-2-SMC from HaCaT cell lysates. Precipitates were resolved on SDS-polyacrylamide gels (10% acrylamide) and probed with SMC2 and SMC4 specific antibodies. SPN, supernatant after pulldown; PD, pulldown. The lower molecular weight band in the SMC2 pulldown is presumably a breakdown product. (b) Immunoprecipitation of SMC2 from HaCaT cell lysates with Nesprin-2 specific mAbK20-478 and of Nesprin-2 with SMC2 specific antibodies. GFP-specific monoclonal antibodies were used for control. The antibodies used for immunoprecipitation are indicated above the panels (IP). The blots were probed with the antibodies listed on the right (WB). Immunoprecipitates were resolved on gradient gels (3–12% acrylamide) and 10% acrylamide gels as appropriate. The data are from one blot; however, the input was not directly adjacent to the SMC2 IP. (c) Interaction of CAP-H2 (condensin II) and CAP-H (condensin I) with Nesprin-2-SMC. Pulldowns were performed with HaCaT cell lysates and GST for control and GST-Nesprin-2-SMC as indicated. Unsynchronized cells were used for the experiments shown in (a)–(c). (d) Analysis of the Nesprin-2-SMC interaction with SMC2 during the cell cycle. HaCaT cells were synchronized with RO-3306 or other reagents as described in Materials and Methods in order to obtain the relevant cell cycle phases. Cell cycle phases were assessed by FACS analysis; the results are depicted in the accompanying diagram. Pulldown was carried out with GST-Nesprin-2-SMC bound to GST-Sepharose. GST was used for control. The blot was probed with SMC2 specific antibodies. (e) Localization of Nesprin-2 as detected with mAb K81-116-6 (green) during mitosis in HaCaT cells. DNA was stained with DAPI. Arrow points to filamentous staining across the chromosomes. (f) Nesprin-2 distribution in HaCaT cells during mitosis as detected with mAb K20-478 (green) and pAbK1 (red). DNA was detected with DAPI. Bar, 10 μm. (g) Nesprin-2 presence on chromosomes. Different Z-stacks (from top to bottom: 0 μm, 0.21 μm 0.42 μm, and 0.84 μm) from a COS7 cell in anaphase stained with mAb K20-478. DNA was stained with DAPI. Bar, 5 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29445399), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Representative human colorectal cancer specimen. Immunohistochemistry of CA9 and GGT1 in mLNs. In these cases, both CA9 and 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 and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30542087), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Modulation of A152T-tau clearance and pathology by upregulation of autophagy. (A and B) Treatment of A152T-tau fish with the autophagy inducers clonidine (A), rapamycin or rilmenidine (B), accelerated clearance kinetics of A152T-tau (mean ± SD of n ≥ 40 neurons/group; Student-Newman-Keuls one-way ANOVA, */#P < 0.05; **/##P < 0.01 ***P < 0.001 versus untreated). (C) Treatment with rapamycin (rap), clonidine (clo) or rilmenidine (ril) also ameliorated morphological defects in A152T-tau transgenic fish (n = 6 independent experiments, 20 fish/group, mean ± SD; Student-Newman-Keuls one-way ANOVA, *P < 0.05; **P < 0.01 and ***P < 0.001 versus DMSO). (D) Representative image showing rapamycin treatment reduced the proportion of abnormal A152T-tau fish. Scale bar = 3.5 mm. (E) Quantification of the escape response measured in individual A152T-tau fish 3 dpf, treated with either DMSO, rapamycin, clonidine or rilmenidine. (n = 5 independent experiments in triplicate, 15/group shown as mean ± SD; two-tailed t-test: *P < 0.05, **P < 0.01 and ***P < 0.001 versus DMSO). The treatment with autophagy upregulators improved the escape response deficit in A152T-tau fish. (F and G) Induction of autophagy by the addition of rapamycin or rilmenidine to A152T-tau fish also reduced levels of phosphorylated tau at residues Ser202/Thr205 (AT8) and Ser396/Ser404 (PHF1) relative to total tau levels (n = 3 independent experiments in duplicate, 10/group shown as mean ± SEM; two-tailed t-test: *P < 0.05 and ***P < 0.001 versus DMSO). (H and I) Treatment with autophagy upregulators results in less cell death. Rilmenidine treatment reduced the levels of active caspase-3 (n = 4 independent experiments in triplicate, 10/group shown as mean ± SEM; two-tailed t-test: *P < 0.05 versus DMSO). (J–N) Injection of an expression vector encoding zebrafish atg5 into A152T-tau fish embryos resulted in over-expression of Atg5 protein at 2 dpf (J and K) (high and low exposure of the same blot presented; mean ± SD, n = 6 independent clutches; two-tailed t-test, *P < 0.05 versus control). (J and L) The increase in Atg5 protein correlated with increase in LC3II, a well-characterized reporter for autophagosome number, demonstrating that autophagy was upregulated in Atg5-injected fish (mean ± SEM, n = 8 independent clutches; two-tailed t-test, ***P < 0.001 versus control). (M and N) A consistent reduction in the number of offspring with morphological defects was observed in Atg5-injected fish compared to control (uninjected) siblings. The percentage of normal fish changes from 52.43% ± 6.01 to 87.79% ± 4.87 after Atg5 injection and consequently, the proportion of abnormal fish diminishes from 47.57% ± 6.01 to 12.21% ± 4.87 (mean ± SEM of seven independent clutches; two-tailed t-test, ***P < 0.001 versus control). Image collected and cropped by CiteAb from the following open publication (//academic.oup.com/brain/article/140/4/1128/2980948), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

SAN1 functions independently of the FA pathway and does not affect FA pathway activation. a, b CSAs of HeLa WT and SAN1−/− cells treated with scrambled ctrl siRNA or FANCD2 siRNA, in response to Cisplatin and MMC (N = 3). Statistical significance determined by two-way ANOVA. Error bars denote s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. c Immunoblot showing siRNA knockdown of FANCD2 in HeLa WT and SAN1−/− cells. d IF staining of FANCD2 foci in HeLa WT cells and SAN1−/− cells treated with 0.045 μM MMC. e Immunoblot of FANCD2 showing mono-ubiquitylation in HeLa WT and SAN1−/− cells treated with vehicle or 0.045 μM MMC. f, g CSAs of HeLa WT and SAN1−/− cells treated with ctrl or SNM1A siRNA and exposed to Cisplatin or MMC. Statistical significance was determined by two-way ANOVA test. h Immunoblot of SNM1A in HeLa WT and SAN1−/− cells treated with ctrl or SNM1A siRNA Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29968717), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Activation of MAPK7 in chondrocytes is essential for hypoxic adaptation and enhancement of HIF1 alpha signaling under hypoxia. a, b Immunofluorescence staining of (a) MAPK7 (b) P-MAPK7 on representative sections of distal femoral growth plates at P1. Scale bar, 200 μm. RZ, resting zone; PZ, proliferative zone; HZ, hypertrophic zone. c Western blot analysis of MAPK7 and p- MAPK7 in primary chondrocytes cultured under hypoxic conditions for 0, 1, 4, 12, or 24 h. d Immunofluorescence staining of SOX9 on representative sections of distal femoral growth plates at P1. Scale bar, 200 μm. e Western blot analysis of SOX9, COL2A1, MAPK7, HIF1 alpha , and VEGFA in CON and Mapk7 CKO chondrocytes cultured under normoxic or hypoxic conditions for 48 h. f Western blot analysis of SOX9, COL2A1, MAPK7, p-MAPK7, HIF1 alpha , and VEGFA protein levels in XMD8-92- or DMSO-treated primary chondrocytes cultured under normoxic or hypoxic conditions. Primary chondrocytes harvested from wild-type mouse knees were cultured in the presence of 5 μM XMD8-92 or an equal amount of DMSO under normoxia for 3 h, followed by normoxia or hypoxia for 48 h. XMD, XMD8-92. g, h mRNA levels of (g) Sox9 and (h) Col2a1 in CON and Mapk7 CKO chondrocytes cultured under normoxia or hypoxia were measured by real-time quantitative PCR (n = 3). *P < 0.05 (one-way ANOVA followed by Dunnett’s post hoc test). Data are presented as mean ± SD. i Immunofluorescence staining of HIF1 alpha on representative proximal tibial growth plate sections from P1 mice. Scale bar, 200 μm. j mRNA levels of Hif1a and its known target genes were measured in growth plate cartilage from P1 Mapk7 CKO mice by real-time quantitative PCR. k HIF1 alpha activity in CON and Mapk7 CKO chondrocytes cultured under normoxia or hypoxia, as evaluated by HIF1 alpha -responsive element luciferase reporter (HRE-luc) assay (n = 5). *P < 0.05 (one-way ANOVA followed by Dunnett’s post hoc test). Data are presented as mean ± SD. l mRNA level of hif1a in CON and Mapk7 CKO chondrocytes cultured under normoxia or hypoxia were measured by real-time quantitative PCR (n = 3). *P < 0.05 (one-way ANOVA followed by Dunnett’s post hoc test). Data are presented as mean ± SD. m Free ATP levels in CON and Mapk7 CKO chondrocytes cultured under normoxic or hypoxic conditions (n = 4). *P < 0.05 (one-way ANOVA followed by Dunnett’s post hoc test). Data are presented as mean ± SD. n Primary chondrocytes were transfected with WT-MAPK7 or DN-MAPK7 expression vector in the presence of CA-MEK5 expression vector. Cells were harvested 48 h after transfection and protein levels of HIF1 alpha , MAPK7 and p-MAPK7 were assayed by western blot Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32944217), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Caspase-11 is expressed in Casp1Null bone marrow-derived macrophages.Immunoblot analysis of caspase-1, caspase-11 and GAPDH (loading control) in unprimed WT or mutant BMDMs at various times (above lane) after stimulation with IFN-beta (250U/ml), IFN-gamma (100 ng/ml) or LPS (100 ng/ml). Data are representative of two independent experiments. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28345580), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

ZNF217 and ALKBH5 regulate NANOG and KLF4 expression via modulation of m6A levelsA. MCF-7 subclones expressing a non-targeting control shRNA (NTC) or shRNAs targeting HIF-1 alpha and HIF-2 alpha (double knockdown [DKD]), were exposed to 20% or 1% O2 for 48 h. Immunoprecipitation of RNA with m6A antibody and RT-qPCR were performed to determine the percentage of KLF4 mRNA with methylation (m6A+) (mean ± SEM; n = 3). **P < 0.01 vs. NTC at 20% O2; ##P < 0.01 vs. NTC at 1% O2 by two-way ANOVA. B. MCF-7 subclones, which were stably transfected with lentiviral vector expressing NTC or either of two independent shRNAs targeting different nucleotide sequences in ZNF217, were exposed to 20% or 1% O2 for 24 h. Total RNA was extracted and m6A levels were determined as a percentage of all adenosine residues (mean ± SEM; n = 3). *P < 0.05, **P < 0.01 vs. NTC at 20%; ##P < 0.01 vs. NTC at 1% O2 by two-way ANOVA. C and D. MCF-7 subclones were exposed to 20% or 1% O2 for 48 h. The percentage of m6A+ NANOG (C) and KLF4 (D) mRNA was determined (mean ± SEM; n = 3). **P < 0.01 vs. NTC at 20% O2; ##P < 0.01, ###P < 0.001 vs. NTC at 1% O2 by two-way ANOVA. E and F. MCF-7 subclones were exposed to 20% or 1% O2 for 24 h. NANOG (E) and KLF4 (F) mRNA levels were determined. *P < 0.05, **P < 0.01, ***P < 0.001 vs. NTC at 20% O2; ##P < 0.01, ###P < 0.001 vs. NTC at 1% O2 by two-way ANOVA. G. MCF-7 subclones were exposed to 20% or 1% O2 for 48 h and immunoblot assays were performed. H. MCF-7 subclones were exposed to 20% or 1% O2 for 48 h. The percentage of m6A+ KLF4 mRNA was determined. *P < 0.05, **P < 0.01 vs. NTC at 20% O2; ##P < 0.01 vs. NTC at 1% O2 by two-way ANOVA. I. MCF-7 subclones were exposed to 20% or 1% O2 for 24 h and KLF4 mRNA levels were determined. *P < 0.05, ***P < 0.001 vs. NTC at 20% O2; ###P < 0.001 vs. NTC at 1% O2 by two-way ANOVA. J. MCF-7 subclones were exposed to 20% or 1% O2 for 48 h and immunoblot assays were performed. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.11743), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

PML-RARa is a HIF-alpha transcriptional co-activator.A HIF-alpha transactivation assays (A–D) with HRE-luciferase construct. Results are presented as Luciferase/Renilla ratio (mean ± s.e.m. of experiments performed in triplicate).B HEK-293 cells transfected with stable mutants of HIF-1 alpha or HIF-2 alpha and increasing concentrations of PML-RAR alpha .C HEK-293 cells transfected with a stable form of HIF-1 alpha along with PML-RAR alpha , PML or RAR alpha .D Wild-type and Pml−/− MEFs transfected with HIF-1 alpha and PML-RAR alpha . Asterisks indicate fold change induction of HIF-1 alpha -mediated transactivation upon PML-RAR alpha expression.E HEK-293 cells transfected with the indicated fusion genes and treated with CoCl2.F Co-immunoprecipitation of exogenous stable forms of HIF-1 alpha (left panel) and HIF-2 alpha (right panel) and PML-RAR alpha with a PML-directed antibody in HEK-293 cells. Of note, exogenously expressed PML-RAR alpha migrates very closely to endogenous PML.Co-immunoprecipitation of endogenous HIF-1 alpha and PML-RAR alpha with a PML-directed antibody in NB4 cells treated with CoCl2.Data information: All experiments were repeated at least twice.Source data are available for this figure. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24711541), 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
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
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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NB100-134

HIF-1 alpha Antibody - BSA Free

(14 Reviews)

(220 Publications)

0.1 ml

NB100-134

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody - BSA Free [NB100-134] - Analysis of HIF-1 alpha in zebrafish retina tissue using NB100-134 HIF-1 alpha antibody. Mouse tissue was used as a control. Image from verified customer review.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - Analysis of HIF-1 alpha in overexpression and endogenous HIF-1 alpha & HIF-2 alpha using anti-HIF-1 alpha antibody NB100-134. The data showed that HIF-1 alpha antibody did not react to HIF-2 alpha overexpression. Image from verified customer review.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - Analysis of HIF-1 alpha using NB100-134, on normoxic and hypoxic nuclear rat cell lysates.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody - BSA Free [NB100-134] - Representative images of macrophages and activated macrophages, in the absence or presence of 10 uM hemin for 72 h. HIF-1 alpha levels were detected by immunofluorescence using a confocal imaging system (Opera Phenix, Perkin Elmer). Images were acquired with 20x water objective. Staining intensity levels in the nucleus and cytosolic region were obtained using Harmony software (Perkin Elmer). Nucleus and cytosol were identified through Hoechst (at 405 nm) and CellMask (at 655 nm) staining, respectively. Scale is shown as 100 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33187129/) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody - BSA Free [NB100-134] - Analysis of HIF-1 alpha in Normoxic (N) and Hypoxic (H) ARPE-19 cells using anti-HIF-1 alpha antibody NB100-134. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody - BSA Free [NB100-134] - Hif1alpha expression in 9.5dpc primordial germ cells. Whole-mount staining of Hif1alpha and GFP in Oct4-GFP embryo (lateral view). Scale bar: 100um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0154309) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody - BSA Free [NB100-134] - Staining in the canine CL on days 10 to 70 after ovulation. PC = positive control (human placenta). NC = negative control. Image from verified customer review.

Immunohistochemistry: HIF-1 alpha Antibody - BSA Free [NB100-134] - Staining of human kidney, renal tubular epithelium in cortex using NB100-134.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody - BSA Free [NB100-134] - Analysis of FFPE tissue section of human endometrium carcinoma AN3CA cell line based xenograft using rabbit polyclonal HIF-1 alpha antibody NB100-134 at 1:300 dilution. The signal was developed using HRP-labelled secondary antibody and DAB reagent, and the section was further counterstained using hematoxylin. The tested section depicted mainly a diffused cytoplasmic staining but there were some cells which showed nuclear signal also (representing hypoxic cells).

Simple Western: HIF-1 alpha Antibody - BSA Free [NB100-134] - Image shows a specific band for HIF-1 alpha in 0.2 mg/mL of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunoprecipitation: HIF-1 alpha Antibody - BSA Free [NB100-134] - PC-3 cell lysates run with cobalt chloride treatments. Immunoprecipitation was performed with protein A/G agarose beads. This data was provided courtesy of Kelie Reece, Figg lab, NCI.

Dual RNAscope ISH-IHC: HIF-1 alpha Antibody - BSA Free [NB100-134] - Formalin-fixed paraffin-embedded tissue sections of human kidney were probed for HIF-1 alpha mRNA (ACD RNAScope Probe, catalog #605228; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog # NB100-134) at 1:500 dilution with one-hour incubation at room temperature 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 cytoplasm of tubules.

Simple Western: HIF-1 alpha Antibody - BSA Free [NB100-134] - Simple Western lane view shows lysates of MCF +/- DMOG, BioSpherix MSCs in hypoxic conditions, and BioSpherix MSCs in normoxic conditions loaded at 0.5 mg/ml. A band was detected for HIF-1 alpha at approximately 116 kDa (as indicated) using NB100-134 (1:100 dilution) followed by Anti-Rabbit Secondary Antibody (042-206, ProteinSimple). This experiment was conducted under standard assay conditions, and using the 12-230 kDa separation module (SW-W004). Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody. Image from an internal validation.

P53 was increased response to IR in the fetal brain.Even though no significant change in protein level, total p53 and phosphorylation of P53 at ser15 were increased compared with HIF-1 alpha in fetal brains response to IR. Numbers in the pictures show the average of three independent experiments (n = 3 from 3 individual pregnant mice).

IGFBP5 inhibits HIF1alpha expression through the MAPK-ERK signaling pathway. C). Overexpression of IGFBP5 inhibited HIF1alpha expression visualized by western blots of A375 cells. The arrow points to the band of HIF1alpha. D. Assessment of the reduced gene expression levels of VEGFA and MMP9, downstream genes regulated by HIF1alpha, in A375 IGFBP5 OE compared to control cells by qRT-PCR analysis. Data were shown for the mean +/- SD form three independent experiments. P values based on two-side Student t-test comparing A375 IGFBP5 OE tumor cells and vector control cells. *, P 25 pictures/group from three separate experiments. Scale bar = 40 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31083380), licensed under a CC-BY licence.

ER-INP binds to PHD2, inhibits hydroxylation of HIF-1alpha and increases HIF-1alpha accumulation. (B) Western blot analysis to measure the effect of ER-INP on HIF-1alpha and its hydroxylation level in transfected HEK293 and RAW264.7 cells pre-treated with MG132 (upper panel) and the protein ratio to beta-actin loading control by ImageJ densitometry analysis (lower panel; n = 3). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31413262), licensed under a CC-BY licence.

HIF-1alpha levels are increased by androgen stimulus and Tip60 overexpression in LNCaP cells. (A) Nuclear and (B) cytosolic HIF-1alpha levels and (C) images in LNCaP cells and in LNCaP cells overexpressing Tip60, in the absence or presence of androgen (10 nM R1881, 72 h). HIF-1alpha levels were detected by immunofluorescence using confocal imaging system. Images were acquired with 20x objective. Staining intensity levels in the nucleus and cytosolic region were obtained using Harmony software. Nucleus and cytosol were identified through Hoechst and CellMask staining, respectively. Scale is shown as 100 um. White dotted frames indicate the section of the image that was enlarged. Values are expressed as mean +/- SEM, from three independent culture preparations, each treatment performed in quadruplicate. Two-way ANOVA, Bonferroni post-test and p values comparisons are specified in the figures (* p < 0.05). HIF-1alpha, hypoxia-inducible factor-1alpha; OE, overexpressing; R1881, synthetic androgen. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31671779), licensed under a CC-BY licence.

The increase of cytosolic HIF-1alpha levels of activated macrophages is inhibited by hemin. (C) Representative images of macrophages and activated macrophages, in the absence or presence of 10 uM hemin for 72 h. HIF-1alpha levels were detected by immunofluorescence using a confocal imaging system (Opera Phenix, Perkin Elmer). Images were acquired with 20x water objective. Staining intensity levels in the nucleus and cytosolic region were obtained using Harmony software (Perkin Elmer). Nucleus and cytosol were identified through Hoechst (at 405 nm) and CellMask (at 655 nm) staining, respectively. Scale is shown as 100 um. White dotted frames indicate the section of the image that was enlarged. Values with different superscript letters (a and b) indicate significant differences (p < 0.05) between groups. For all bars with the same letter, the difference between the means is not statistically significant. HIF-1alpha, hypoxia-inducible factor-1 alpha. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33187129), licensed under a CC-BY licence.

LOXL2 abundance in EC-derived exosomes is increased in hypoxia. (A) Immunoblots of endothelial cells for HIF-1alpha (control for hypoxia, top panel), LOXL2 (middle panel) and beta-actin (loading control, lower panel). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26612622), licensed under a CC-BY licence.

Hypoxic setup for larval zebrafish. (D) Western blot of Hif1alpha protein expression showing that hypoxia enhanced Mbp protein translation. *P < 0.05; **P < 0.01; ***P < 0.001. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30337858), licensed under a CC-BY licence.

Enhanced expression of hypoxia-induced survival factors and angiogenic growth factors in hASC L-spheroids. (C) Western blot analysis and quantification of HIF1-alpha in hASCs cultured as spheroids, L-spheroids and monolayers (*p < 0.01, compared to the L-spheroid group). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26065900), licensed under a CC-BY licence.

E2 enhances neutrophil apoptosis through suppressing the expression of hif1alpha and c-myb under physiological conditions. C) Staining of hif1alpha with antibody with or without E2 treatment. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35842445), licensed under a CC-BY licence.

HIF1alpha and AhR mediate butyrate induction of IL-22 in CD4+ T cells.. b-f CD4+ T cells were activated with anti-CD3/CD28 mAbs +/- butyrate (0.5 mM) under Th1 conditions (n = 3/group). Hif1a (b) and Ahr (c) were analyzed by qRT-PCR. HIF1alpha (d) and AhR (e) protein was analyzed by western blot on day 2. HIF1alpha activity was measured using HIF1alpha Transcription Factor Assay Kit (f). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32901017), licensed under a CC-BY licence.

$Alteration of PARKIN leads to an impairment of mitophagy upon CCCP treatment.a 2 N and DS were treated with CCCP (20 μM) for 6 h and mitochondrion/cytosol fractionation was performed to assay the accumulation of LC3-II and p62 within the mitochondrial fraction. VDAC1 and ACTIN were used as a loading control for mitochondrial and cytosolic fractions, respectively. b, c Quantitative data of LC3-II and p62 in the mitochondrial fraction were normalized with VDAC1 (n = 3). d The cells were treated with CCCP (20 μM) for 6 h and the whole-cell extracts from 2 N and DS were analyzed by western blot for PARKIN and PINK1. e Quantification of PARKIN normalized with ACTIN (n ≥ 3). d Quantification of PINK1 after 6 h of CCCP, each quantitative data was normalized with ACTIN (n ≥ 3). g Representative confocal images of cells stained with Mitotracker green (mitochondrial marker) and Lysotracker red (lysosomal marker) upon CCCP treatment (20 μM for 6 h). White arrow heads point to the co-localization within puncta from the boxed region at high magnification shown on the right. Scale bar 10 μm. h The graph shows the percentage of Mitotracker green /Lysotracker red colocalization calculated by JACoP plugin of ImageJ. Minimum 50 cells per condition were counted from three independent experiments. i 2 N and DS cells were treated with CCCP (20 μM) for 6 h and were assessed for SQSTM1 (p62), PARKIN and PINK1 mRNA by quantitative real-time PCR. mRNA levels were normalized to ACTIN mRNA, used as internal control. Data display the fold-changes of SQSTM1, PARKIN and PINK1 mRNAs relative to control cells (n = 3). Statistical analysis was performed using Student’s T-test or one-way ANOVA with Tukey’s multiple comparisons test. (*p < 0.05; **p < 0.01; ***p < 0.001; n.s. no significant difference). Immunoblots reported are from one representative experiment Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31332166), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Metformin synergized with sorafenib to suppress angiogenesis, proliferation, and EMT but promote apoptosis in vivo. Representative images of tumor sections were stained with antibody against CD31, Ki67, and TUNEL (magnification ×200). a Metformin in combination with sorafenib synergistically minimized both Ki67 and CD31 expression but promoted TUNEL expression. b Combined therapy inhibited EMT at protein level which lysated from the tumor tissue, and pAMPK expression was elevated both in metformin group and combined treatment groups. c Combined treatment of metformin and sorafenib suppressed HIF-2 alpha protein expression but upregulated TIP30 protein expression evaluated by Western blot Image collected and cropped by CiteAb from the following open publication (//www.jhoonline.org/content/9/1/20), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

EBNA1 expression is associated with upregulation of MTH1 and oxidative damage repair pathways. The expression of MTH1, OGG1, and MUTYH was investigated by western blots and qPCR in BJAB-tTAE1 cells upon addition and withdrawal of doxycycline. a Representative western blots illustrating the correlation between the reversible down- and upregulation of MTH1 and EBNA1 in BJAB-tTAE1 cells upon addition and withdrawal of doxycycline. GAPDH was used as loading control. b Densitometry quantification of MTH1 and EBNA1 expression in BJAB-tTAE1 cells. The mean intensity of the MTH1 and EBNA1 specific bands relative to GAPDH in three independent experiments is shown in the figure. c Regression analysis of the relationship between expression levels of MTH1 and EBNA1. The data from three independent experiments were used for the plot. d Representative western blots illustrating the expression of MTH1, MUTYH, and OGG1 in BJAB-tTAE1 cells cultured for two weeks in the presence or absence of doxycycline. e Fold change is the ratio between the intensity of the specific band in cells cultured without or with doxycycline. The mean ± SD of four independent experiments is shown. f qPCR analysis of the levels of MTH1, OGG1, and MUTYH transcripts in BJAB-tTAE1 cells cultured for 2 weeks in the presence or absence of doxycycline. The mean ± SE of the fold change in six independent experiments each performed in triplicate is shown Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31511648), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Impacts of PRDX3 protein on autophagy flux(A–D) Representative immunoblot (A, C) and quantification (B, D) showing the levels of LC3-II in BPH-1 cells treated with random (MOCK) or PRDX3-specific siRNA (PRDX3) (A, B) or RWPE-1 cells transiently expressing different amount of PRDX3 (C, D) in the absence (Ctrl) or presence of bafilomycin A1 (BAF). Data are mean and standard deviation of three repeats and differences are tested with Student's T-test. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001. (E–G) Representative immunoblot (E) and quantification (F, G) showing the levels of Beclin 1 (F) and PI3KCIII (G) in BPH-1 cells treated with random (MOCK) or PRDX3-specific siRNA (PRDX3). Ns, not significant; *P ≤ 0.05. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17927), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Autophagy level was promoted by downregulation of the Hh signaling pathway in zebrafish larvae. (A–C) Larvae were exposed to 10 µM cyA and 0.1% DMSO at 3 dpf for 3 days. (D–H) Embryos were injected with SHHMO, Gli2MO and Gli2Mis at the 1–4 cell stage; at 6 dpf, larvae were collected for assays . GAPDH served as a loading control. The means and s.d. were derived from triple duplicates. *P<0.05, **P<0.01, ***P<0.001 versus the WT groups or DMSO control groups (one-way ANOVA). The data were from three independent experiments. (A,D) SHH protein was detected by western blot assay. (B,E,G) Transcriptional level of autophagy-related genes was tested by qPCR assays. (C,F,H) Autophagy-related proteins LC3B II, ATG5 and P62 level were determined by western blot. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30992325), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

FASN Regulates HIF-alpha Protein Stability and HIF Target Gene Expression. (A) FASN siRNA downregulates HIF-alpha protein levels. PREC, HeLa and HCT116 cells were treated with FASN (+) or control (−) siRNA and harvested after 72 h. Cells were exposed to hypoxia (1% oxygen) or maintained in normoxia (21% oxygen) 4 h before harvesting and subjecting WCEs to western blot analysis. Unmodified (250 kDa) and modified (larger) FASN bands are indicated by large arrowheads, REDD1 by a small arrowhead, and HIF-1 alpha and HIF-2 alpha by arrows. (B) Downregulation of HIF-1 alpha and HIF-2 alpha in HCT116 cell clones carrying the FASN gene mutagenized using CRISPR-Cas9 (See Supplementary Fig. S3 for genomic DNA information). Protein samples were prepared in normoxia (left column) or hypoxia (right column) for western blot analysis. Asterisk indicates a non-specific band. (C) Quantification of HIF-target gene expression by qPCR upon downregulation of FASN in PREC. Samples were prepared from cells treated as described for panel A but with 24 h hypoxia. siRNAs were transfected in triplicates and mRNAs of the HIF target genes were measured in biological triplicates and technical duplicates as a total of 6 samples by qPCR. Values were normalized against the PPIA reference gene and control samples. (D) Loss of downregulation of HIF-2 alpha levels in ccRCC cells. pVHL-deficient 786-O and A498 cells were tested in FASN siRNA knockdown experiments. (E) Restoration of HIF-alpha regulation by stable expression of exogenous pVHL in SKRC10 cells. pVHL-WT or pVHL-Y112H were introduced into pVHL-deficient SKRC10 ccRCCs using a lentiviral vector carrying a weak promoter and HIF-alpha protein levels were examined. Specific and non-specific bands are indicated by arrows and an asterisk, respectively. (F) FASN and the E3 ligase components are present in the pVHL-containing complexes in a mutually exclusive manner. The indicated constructs were transfected into 293 T cells and WCEs prepared after 48 h were tested for protein-protein interactions by precipitation with an anti-Flag antibody-conjugated resin. Note that FASN failed to precipitate CUL2 and Elongin C, as shown in lanes 6 and 8. Experiments were repeated twice (D,E,F) or more (A,B), and the results were reproduced. The qPCR experiment (C) was performed once with biological triplicates. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28775317), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Etv5b activates the smn promoter leading to increased Smn levels in motoneurons.(a–d) Whole-mount in situ hybridization of etv5b expression in 24 hpf embryos. Expression pattern in the entire embryo (a), in magnified lateral (b) and dorsal (c) views as well as in a transverse section (d) of the trunk. In the latter case, yellow arrowhead denote signal in the motoneuron region. Scale bars: 100 μm (a) and 50 μm (b–d). nc, notochord. (e) RT-PCR in control (C - only psmn:mCherry) versus etv5b overexpressing (OE - etv5b mRNA and psmn:mCherry) embryos at early gastrula stage. (f,g) scIF of control (f) and etv5b MO (g) cells from pmnx1:eGFP embryos. H2B, GFP and Smn signals are shown in Z-projected confocal sections. GFP+ cells marked by white rectangles are magnified on the right. Scale bars: 10 μm for low and 5 μm for high magnification. (h) Relative Smn signal in control and in etv5b MO cells from one representative experiment. Exact values are (mean ± SD): 0.99 ± 0.25 (control) and 0.7 ± 0.14 (etvb5 MO), p = 0.003 with Wilcoxon Sum Rank Test. (i) Average relative Smn levels in control and in etv5b MO motoneurons. The exact value of decrease is 0.75 ± 0.04 (mean ± SD). N = 3 experiments, n = number of analyzed cells. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/srep27470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

The A2AR and baicalin attenuated CXCR4 expression in the hypoxia-induced PAH mouse model. CXCR4 and MYH11 expression levels in mouse PASMCs were analyzed by immunofluorescence staining (a; n = 3). CXCR4 protein is stained red, and MYH11 is stained green to indicate the PASMCs (×400; scale bars indicate 50 μm). CXCR4 fluorescence intensity was calculated (b; n = 3). CXCR4 protein expression levels in lung tissue were examined by western blot. GAPDH served as an internal control (c, d; n = 3). Data are presented as the mean ± SD. #Value significantly greater than the corresponding value in saline-treated normoxic mice (##P < 0.01). *Value significantly less than the corresponding value in hypoxic mice (*P < 0.05, **P < 0.01). §Value significantly less than the corresponding value in baicalin-treated mice (§P < 0.05, §§P < 0.01). +Value significantly different between WT and A2AR−/− mice (+P < 0.05, ++P < 0.01). WTH, wild-type hypoxic; A2AR−/−H, A2AR−/− hypoxic; s, saline-treated Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28774332), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bv, Ca, Gp, Pm, Xp, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Bv, Ca, Gp, Pm, Xp, Ze

Applications:

, , , , , , , , , ,

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

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-134AF350
Alexa Fluor 405	NB100-134AF405
Alexa Fluor 488	NB100-134AF488
Alexa Fluor 532	NB100-134AF532
Alexa Fluor 594	NB100-134AF594
Alexa Fluor 647	NB100-134AF647
Alexa Fluor 700	NB100-134AF700
Alexa Fluor 750	NB100-134AF750
Allophycocyanin	NB100-134APC
Biotin	NB100-134B
DyLight 350	NB100-134UV
DyLight 405	NB100-134V
DyLight 488	NB100-134G
DyLight 550	NB100-134R
DyLight 594	NB100-134DL594
DyLight 650	NB100-134C
DyLight 680	NB100-134FR
DyLight 755	NB100-134IR
FITC	NB100-134F
HRP	NB100-134H
Janelia Fluor 549	NB100-134JF549
Janelia Fluor 646	NB100-134JF646
PE	NB100-134PE
PerCP	NB100-134PCP
mFluor Violet 450 SE	NB100-134MFV450
mFluor Violet 500 SE	NB100-134MFV500
mFluor Violet 610 SE	NB100-134MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-134SS

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NB100-690

alpha Tubulin Antibody (DM1A) - BSA Free

(14 Reviews)

(134 Publications)

0.1 ml

NB100-690

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

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

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

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

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

Western Blot: alpha Tubulin Antibody (DM1A) [NB100-690] - GS treatment increases markers of beiging in 3T3-L1 adipocytes. GS treatment upregulates markers of beiging, including beta-3AR (C) proteins. Data presented as mean +/- SEM from n = 4 replicates per group. * p < 0.05, *** p < 0.001 vs. control. Abbreviations: beta-3 adrenergic receptor (beta-3AR). Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2305-6320/6/1/22), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) - BSA Free [NB100-690] - Mouse MS1 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 alpha Tubulin Antibody [DM1A] conjugated to Alexa Fluor 647 (NB100-690AF647) at 2 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunohistochemistry-Paraffin: alpha Tubulin Antibody (DM1A) [NB100-690] - IHC analysis of a formalin fixed and paraffin embedded tissue section of mouse prostate using alpha Tubulin Antibody (DM1A) at 1:200 dilution. The signal was developed using HRP labelled secondary and DAB reagent which followed counterstaining with hematoxylin. The antibody generated a specific cytoplasmic/cytoskeletal staining in the prostate epithelial cells.

Flow Cytometry: alpha Tubulin Antibody (DM1A) [NB100-690] - An intracellular stain was performed on HeLa cells with alpha Tubulin [DM1A] Antibody NB100-690AF700 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 700.

Western Blot: alpha Tubulin Antibody (DM1A) [NB100-690] - Western blot analysis of extracts from HeLa, COS and C6 cells using alpha Tubulin antibody (NB100-690, 1:1000, Alpha tubulin molecular weight: 50 kDa)

Western Blot: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of alpha tubulin (molecular weight of 50 kDa) in 9 cell lysates. Lane 1. HeLa; Lane 2. JURKAT; Lane 3. COS7; Lane 4. NIH-3T3; Lane 5. PC-12; Lane 6. RAT2; Lane 7. CHO; Lane 8. MDBK; Lane 9. MDCK

Western Blot: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of HeLa and COS-7 lysates. Alpha tubulin molecular weight: 50 kDa.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - IF Confocal analysis of C6 cells using alpha Tubulin antibody (NB100-690, 1:50). An Alexa Fluor 488-conjugated Goat to mouse IgG was used as secondary antibody (green, A). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red, B). DAPI was used to stain the cell nuclei (blue, C).

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-alpha Tubulin (DM1A) (NB100-690) at a 1:200 dilution overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - Staining of skin fibroblasts.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of embryonic fibroblasts in the anaphase portion of mitosis.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - U-251 MG 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-alpha Tubulin Antibody [DM1A] conjugated to Alexa Fluor 488 (NB100-690AF488) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - 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-alpha Tubulin Antibody [DM1A] conjugated to Alexa Fluor 488 (NB100-690AF488) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - NIH3T3 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-alpha Tubulin Antibody [DM1A] conjugated to Alexa Fluor 488 (NB100-690AF488) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - HeLa 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 alpha Tubulin Antibody [DM1A] conjugated to Janelia Fluor 549 (NB100-690JF549) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunohistochemistry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of paraffin embedded colon sections.

Immunohistochemistry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of small intestine tissue fixed with formalin and paraffin embedded showing cytoplasmic and cytoskeletal staining of glandular cells.

Immunohistochemistry-Paraffin: alpha Tubulin Antibody (DM1A) [NB100-690] - IHC analysis of a formalin fixed paraffin embedded tissue section of mouse skeletal muscle using alpha Tubulin Antibody (DM1A) at 1:100 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a strong cytoplasmic signal in the muscle cells with cytoplasmic-nuclear signal in the endothelial cells.

Flow Cytometry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of PE conjugate of NB100-690. An intracellular stain was performed on RAW 246.7 cells with Alpha Tubulin antibody (DM1A) NB100-690PE (blue) and a matched isotype control NBP2-27287PE (orange). Cells were fixed with 4% PFA and then permeablized wi

Flow Cytometry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of PE conjugate of NB100-690. An intracellular stain was performed on SH-SY5Y cells with Alpha Tubulin antibody (DM1A) NB100-690PE (blue) and a matched isotype control NBP2-27287PE (orange). Cells were fixed with 4% PFA and then permeablized with

Flow (Intracellular): alpha Tubulin Antibody (DM1A) [NB100-690] - An intracellular stain was performed on HeLa cells with alpha Tubulin Antibody (DM1A) NB100-690AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488. Image from the Alexa Fluor 488 version of this antibody.

Immunomicroscopy: alpha Tubulin Antibody (DM1A) [NB100-690] - Staining of the marine parasite Cryptocaryon irritans mouth. Large bundles of microtubules form a cytophyrigeal basket.

GS treatment increases markers of beiging in 3T3-L1 adipocytes. GS treatment upregulates markers of beiging, including UCP1 (A), TBX1 (B), and beta -3AR (C) proteins. Data presented as mean ± SEM from n = 4 replicates per group. * p < 0.05, *** p < 0.001 vs. control. Abbreviations: isoproterenol (ISO), uncoupling protein 1 (UCP1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), T-box protein 1 (TBX1), beta -3 adrenergic receptor ( beta -3AR).

Simvastatin increases cytotoxicity in lung cancer cells. (F) Western blots of indicated proteins involved in apoptosis and autophagy in both Bm7 and HCC827 cells with control (shGFP) and p53 knockdown (shTP53) treated with simvastatin is shown. MDM2, murine double minute 2; AKT, serine-threonine kinase; PARP, poly (ADP-ribose) polymerase; mTOR, mammalian target of rapamycin; WT, wild type. Full-length blots/gels are presented in Supplementary Fig. 1. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31892709), licensed under a CC-BY licence.

$Role of HIF-1 alpha , FOXO3 in hypoxia-induced BNIP3 expression. (A) UCB-hMSCs were incubated with various durations of hypoxia (0–48 h). The protein expressions of HIF-1 alpha and beta -actin were detected by western blot. n = 4. (B) UCB-hMSCs were pretreated with NAC (5 mM) for 30 min prior to hypoxia incubation for 24 h. The protein expressions of HIF-1 alpha , lamin A/C and beta -tubulin in non-nuclear and nuclear fractionized cell samples were assessed by using western blot. n = 3. (C) UCB-hMSCs were immuno-stained with HIF-1 alpha and PI (magnification ×600). Scale bars, 37.5 µm. (D) HIF1A siRNA or NT siRNA was transfected to cells prior to hypoxia treatment for 24 h. The mRNA expression of BNIP3 was analyzed by qPCR. n = 6. (E) The protein expressions of BNIP3 and HIF-1 alpha were detected by western blot. n = 4. (F) NAC (5 mM) was pretreated to UCB-hMSCs prior to hypoxia treatment for 24 h. FOXO3, lamin A/C and beta -tubulin proteins expressions were assessed by western blot. n = 3. (G) FOXO3 siRNA transfected to UCB-hMSCs prior to hypoxia treatment for 24 h. The FOXO3 mRNA expression was measured by qPCR. n = 6. (H) BNIP3, FOXO3 and beta -actin expressions were detected by western blot. n = 3. Western blot data were normalized by beta -actin, and qPCR data were normalized by ACTB mRNA expression level. Lamin A/C and beta -tubulin were used as nuclear and non-nuclear protein controls, respectively. Quantitative data are presented as a mean ± S.E.M. All blots and confocal images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected and cropped by CiteAb from the following open publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

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

Alpha Tubulin (DM1A) was detected in immersion fixed U-251 MG human glioblastoma cell line using Mouse anti-alpha Tubulin (DM1A) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor® 647 (Catalog # NB100-690AF647) (light blue) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (dark blue). Cells were imaged using 100X objective and digitally deconvolved.

Reactivity:

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

Species Glossary

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

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

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

Available Formulations

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

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NB300-146

Sodium Potassium ATPase Alpha 1 Antibody (464.6)

(4 Reviews)

(114 Publications)

0.05 ml

NB300-146

Immunohistochemistry: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - Staining of human enodmetrial glands within the uterus using NB300-146. Note the absence of staining in the surrounding myometrial smooth muscle.

Flow Cytometry: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - An intracellular stain was performed on NIH3T3 cells with Sodium Potassium ATPase Alpha 1 Antibody (464.6) NB300-146 (blue) and a matched isotype control MAB002 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (84540, Thermo Fisher).

Western Blot: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - Analysis detecting Na, K-ATPase (alpha) in porcine proximal tubule protein.

Immunocytochemistry/Immunofluorescence: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - Hek293 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-Sodium Potassium ATPase Alpha 1 (464.6) at 10 ug/ml overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/Immunofluorescence: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-Sodium Potassium ATPase Alpha 1 (464.6) at 10 ug/ml overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/Immunofluorescence: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - Detection of ATPA1 (Green) in HepG2 cells using NB300-146. Nuclei (Blue) were counterstained using Hoechst 33258.

Immunocytochemistry/Immunofluorescence: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - ARPE-19 cells cultured on transwell inserts for 4 weeks are not completely polarized. ARPE-19 cells were cultured up to 4 weeks on transwell inserts covered with laminin. The expression of Na+, K+-ATPase using anti-alpha1 antibody was observed mostly at the basolateral membrane. Scale bar: 10 um. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fphys.2016.00450/full), licensed under a CC-BY license.

Flow Cytometry: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - An intracellular stain was performed on A549 cells with NB300-146 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 1 ug/mL for 30 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems).

Flow Cytometry: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - An intracellular stain was performed on A549 cells with NBP2-61137AF647 (Blue) and a matched isotype control (Orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647. Image using the Alexa Fluor 647 form of this antibody.

Flow Cytometry: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - An intracellular stain was performed on A549 cells with - Purified NBP2-61137 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 1 ug/mL for 30 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems). Image using the Purified form of this antibody.

Flow Cytometry: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - An intracellular stain was performed on RCC4 cells with Sodium Potassium ATPase Alpha 1 Antibody (464.6) NB300-146 (blue) and a matched isotype control MAB002 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (84540, Thermo Fisher).

Flow Cytometry: Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - An intracellular stain was performed on SK-MEL-28 cells with Sodium Potassium ATPase Alpha 1 Antibody (464.6) NB300-146 (blue) and a matched isotype control MAB002 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (84540, Thermo Fisher).

Sodium Potassium ATPase Alpha 1 Antibody (464.6) [NB300-146] - ARPE-19 cells cultured on transwell inserts for 4 weeks are not completely polarized. ARPE-19 cells were cultured up to 4 weeks on transwell inserts covered with laminin. The expression of Na+, K+-ATPase using anti-alpha1 antibody was observed mostly at the basolateral membrane. Scale bar: 10 um. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fphys.2016.00450/full), licensed under a CC-BY license.

SDS-PAGE western blot validation of the increased abundance of Annexin V, Na/K ATPase and alpha-tubulin in the EVs derived from TIS Cal51 cells compared with EVs derived from non-senescent non-treated controls. EVs isolated from TIS and control cells were immunoblotted to validate the protein expression of annexin V, Na/K ATPase and alpha-tubulin from the MS data. CD63 and HSP70 were used as loading controls. Densitometry was completed using Image J software (Fiji). All values are expressed as the mean of three independent experiments+/-s.e.m. The limit of +/-1 skewness was set as normal distribution. Skewed data were transformed to Y=Log(Y) to fit a normal distribution. Results were analysed using the Student's t-test with significant differences having a *P<=0.05, **P<=0.01, ***P<=0.001. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/oncsis201782), licensed under a CC-BY licence.

Transfected SK-N-AS cells express ICAM-2 transcripts and proteins. F) ICAM-2 WT and variants localized to cell membranes. Experimental details are included in Methods. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23714211), licensed under a CC-BY licence.

ARPE-19 cells cultured on transwell inserts for 4 weeks are not completely polarized. ARPE-19 cells were cultured up to 4 weeks on transwell inserts covered with laminin. Th The expression of Na+, K+-ATPase using anti-alpha1(C) and anti-beta1 antibodies (D) was observed mostly at the basolateral membrane. Immunofluorescence detection with anti-human beta2 antibody revealed a very weak signal (E). Scale bar: 10 um. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fphys.2016.00450/full), licensed under a CC-BY licence.

Endothelial-mesenchymal transformation (EMT) of RCECs using specific endothelial-mesenchymal transformation medium (SEMTM). (C) Western blots showed that Fresh RCECs and EMT-RCECs expressed Vimentin, Atp1a1, a marker of differentiated endothelial cells, and b-actin. Original gel and blots are shown in Supplemental Figs S3, 4. Semiquantitative analysis shows significant upregulation of the protein levels of Vimentin (p = 0.048) and downregulation of the protein level of Atp1a1 (p = 0.0043). Data expressed as mean +/- SD of three replicate experiments. Student's t test (*p < 0.05, **p < 0.01). (n = 3). RCECs: rabbit corneal endothelial cells, alpha-SMA: smooth muscle alpha-actin. Atp1a1: Na,K-ATPase alpha-subunit. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30442918), licensed under a CC-BY licence.

The cellular model of the blood-CSF barrier. (D) Immunofluorescent staining of Na+K+ ATPase and ABCC1, showing the expected respective apical and basolateral membrane localization in the choroidal epithelial cells. The left image is a close up of a single cell to better appreciate the polarity of distribution of the 2 proteins. Nuclei appear in blue. Arrows show the lateral cellular membranes best seen in the z direction by confocal analysis, arrowheads show the basal labeling of ABCC1. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0150945), licensed under a CC-BY licence.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ca, Dr, Gp, Pm, Rb, Sh, Xp, Ye

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Dr, Gp, Pm, Rb, Sh, Xp, Ye

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-61137AF350
Alexa Fluor 405	NBP2-61137AF405
Alexa Fluor 488	NBP2-61137AF488
Alexa Fluor 532	NBP2-61137AF532
Alexa Fluor 594	NBP2-61137AF594
Alexa Fluor 647	NBP2-61137AF647
Alexa Fluor 700	NBP2-61137AF700
Alexa Fluor 750	NBP2-61137AF750
Allophycocyanin	NBP2-61137APC
Biotin	NBP2-61137B
DyLight 350	NBP2-61137UV
DyLight 405	NBP2-61137V
DyLight 488	NBP2-61137G
DyLight 550	NBP2-61137R
DyLight 594	NBP2-61137DL594
DyLight 650	NBP2-61137C
DyLight 680	NBP2-61137FR
DyLight 755	NBP2-61137IR
FITC	NBP2-61137F
HRP	NBP2-61137H
Janelia Fluor 549	NBP2-61137JF549
Janelia Fluor 646	NBP2-61137JF646
PE	NBP2-61137PE
PerCP	NBP2-61137PCP
mFluor Violet 450 SE	NBP2-61137MFV450
mFluor Violet 500 SE	NBP2-61137MFV500
mFluor Violet 610 SE	NBP2-61137MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-61137
Azide and BSA Free	NBP2-80956

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NB100-355

RPE65 Antibody (401.8B11.3D9) - BSA Free

(7 Reviews)

(93 Publications)

0.2 ml

NB100-355

Immunohistochemistry-Paraffin: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Analysis of FFPE human glioblastoma tissue section using 1:500 dilution of RPE65 [401.8B11.3D9] antibody on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems).

$Western Blot: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - WB analysis of RPE65 in 20ug lysate of COS7 cells expressing recombinant Human RPE65 (Lane 1) and 5ug of Bovine retinal pigment epithelium membrane fraction (Lane 2). Blot processed for detection with alkaline phosphatase conjugated goat-anti Mouse IgG secondary antibody and NBT/BCIP substrate.$

Immunocytochemistry/Immunofluorescence: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - ICC-IF analysis of cultured ARPE19 cells, a spontaneously arising human retinal pigment epithelia cell line - 10 minutes fixation in 4% PFA, 10 minutes permeabilization in PBS containing 0.2% Triton X-100 (PBS-T), 1 hour blocking in 10% normal goat serum containing 1% BSA in PBS-T, 1:100 primary antibody dilution in PBS, ON 4C incubation.

Immunohistochemistry-Frozen: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Staining of RPE65 in a cryosection of mouse retina tissue using RPE65 antibody (clone 401.8B11.3D9).

Immunohistochemistry: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Zebrafish brain ventricular area labeled with mouse anti-RPE65 (1:100). This image was submitted through a verified customer review.

Simple Western: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - iPSC-RPE from all donors express RPE marker proteins. iPSC-RPE from the patient, the patients mom, the patients dad, and the three unrelated, unaffected controls all expressed RPE65. The negative control (iPSCs) had no detectable RPE65 protein expression. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29540715/) licensed under a CC-BY license.

Knockout Validated: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Immunoblotting showing the expression of RPE-specific proteins BEST1 (NB300-164), RPE65 (NB100-355), CRALBP, and the loading control beta-Actin in iPSC-RPE cells. Two gels/blots in the same panel were prepared from the same cell lysate of each PSC-RPE to detect BEST1 + beta-Actin, and RPE65 + CRALBP, respectively. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34061021/) licensed under a CC-BY license.

Attenuation of mTORC1/2 kinase ablates CPT-induced p53- and p53-dependent responses in ccRCC. (a) The 786-O and 786-O/VHL cells were treated with 400 nM pp242, 2 μM CPT, 10 μM nutlin-3a (N3) or vehicle control (DMSO) alone for 24 h, or preincubated with 400 nM pp242 for 1 h before addition of 2 μM CPT or 10 μM N3 for a further 24 h. Whole-cell lysates were assayed by western blot for HIF-2 alpha , mTOR, phosphorylated p53 (S15), mTOR (S2448) and p70S6K (T389) proteins. Actin was used as a loading control. Short and long exposure times are shown for P-S15-p53. (b) RCC4 and RCC4/VHL cells were treated with 400 nM pp242 or vehicle control (DMSO) alone or preincubated with 400 nM pp242 for 1 h before addition of either 1 μM or 2 μM CPT for 24 h. Whole-cell lysates were assayed by western blot for mTOR, p53, phosphorylated p53 (S15), mTOR (S2448) and p70S6K (T389) proteins. Actin was used as a loading control. (c) RCC4 and RCC4/VHL cells were preincubated with 400 nM pp242 for 1 h before addition of 2 μM CPT for 24 h and mRNA expression of ET-1 and PAI-1 were assessed by real-time quantitative PCR relative to GAPDH. Mean±S.E. of duplicate values of one representative experiment is shown. (d) RCC4 cells were pretreated with 400 nM pp242 for 1 h before addition of 2 μM CPT for 24 h. Conditioned media were harvested and assessed for secreted ET-1 protein levels by ELISA and normalized to total protein levels. Mean±S.E. of duplicate values of one representative experiment is shown Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24136229), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Depletion of TCAB1 reduced the cell proliferation in vitro. a. qPCR results. Using exogenous shTCAB1 lentivirus (4 different shRNA targets) depleted TCAB1 mRNA significantly. b. Protein level also significantly decreased after shTCAB1 lentivirus treatment. c. Depletion of TCAB1 by exogenous shRNA decreased cell proliferation in Cal-27, ACC2 and HSC-3 cells. d. FACS results. Depletion of TCAB1 might facilitate cancer cell arrest, and shTCAB1-treated cells are significantly less in S phase. Statistical analysis was determined by Student’s t test (**P < 0.01, ***P < 0.005). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25070141), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Cortisol inhibited selective autophagy towards SCG10 via activation of mTOR.a The cells were treated with cortisol (1 μM) for 3 h, and stained with rhod-2 (3 μM) for 1 h to detect mitochondrial Ca2+. After incubation, mitotracker green (MTG, 300 nM) was also stained to visualize mitochondria. The intensity of both rhod-2 (red) and MTG (green) was measured with Eclipse Ts2™ fluorescence microscopy. ** indicates p < 0.01 vs. control. Scale bars represent 100 μm (magnification, ×400). n = 5. b The cells were treated with RU 486 (1 μM) for 30 min before cortisol (1 μM) for 6 h, and then reacted with ATP luciferase reagent. The ATP levels were detected with luminometer. ** indicates p < 0.01 vs. control and ## indicates p < 0.01 vs. cortisol alone. n = 6. c Time responses (0–6 h) of cortisol (1 μM) in phosphorylation of AMPK at Thr172 and mTOR at Ser2448 were shown. ** indicates p < 0.01 vs. control. n = 4. d The cells were treated with xestospongin C (1 μM) for 2 h or ruthenium red (100 nM) for 30 min before cortisol (1 μM) for 6 h. p-mTOR (Ser2448), mTOR, and beta -actin were detected in western blotting results. * indicates p < 0.05 vs. control and # indicates p < 0.05 vs. cortisol alone. n = 4. e The cells were pretreated with rapamycin (100 nM) for 30 min before cortisol (1 μM) for 24 h. Atg5, p62, LC3, and beta -actin were detected with western blot. ** indicates p < 0.01 vs. control. #, ## indicates p < 0.05, p < 0.01 vs. cortisol, respectively. n = 5. f Time responses (0–24 h) of cortisol (1 μM) in stathmin-1 and SCG10 expressions were shown. *, ** indicates p < 0.05, p < 0.01 vs. control, respectively. n = 5. g The cells were pretreated with rapamycin (100 nM) for 30 min before cortisol (1 μM) for 24 h. SCG10 and beta -actin were detected with western blot. * indicates p < 0.05 vs. control and ## indicates p < 0.01 vs. cortisol. n = 4. h The cells were pretreated with rapamycin (100 nM) for 30 min before cortisol (1 μM) for 24 h. SCG10 was co-immunoprecipitated with an anti-ubiquitin antibody (the left side). Expression of ubiquitin, SCG10, and beta -actin in total cell lysates is shown in the right side. ** indicates p < 0.01 vs. control and ## indicates p < 0.01 vs. cortisol alone. n = 4. i The cells were pretreated with rapamycin (100 nM) for 30 min before cortisol (1 μM) for 24 h. Co-localization of LC3 (green) and SCG10 (red) was visualized with SRRF imaging system. DAPI was used for nuclear counterstaining (blue). ** indicates p < 0.01 vs. control and ## indicates p < 0.01 vs. cortisol alone. Scale bars represent 20 μm (magnification, ×1,000). n = 5. All blot and immunofluorescence images are representative. Quantative data are presented as a mean ± S.E.M. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30429451), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ca, Ch, Pm, Xp

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Ch, Pm, Xp

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-355AF350
Alexa Fluor 405	NB100-355AF405
Alexa Fluor 488	NB100-355AF488
Alexa Fluor 532	NB100-355AF532
Alexa Fluor 594	NB100-355AF594
Alexa Fluor 647	NB100-355AF647
Alexa Fluor 700	NB100-355AF700
Alexa Fluor 750	NB100-355AF750
Allophycocyanin	NB100-355APC
Biotin	NB100-355B
DyLight 350	NB100-355UV
DyLight 405	NB100-355V
DyLight 488	NB100-355G
DyLight 550	NB100-355R
DyLight 594	NB100-355DL594
DyLight 650	NB100-355C
DyLight 680	NB100-355FR
DyLight 755	NB100-355IR
FITC	NB100-355F
HRP	NB100-355H
Janelia Fluor 549	NB100-355JF549
Janelia Fluor 646	NB100-355JF646
PE	NB100-355PE
PerCP	NB100-355PCP

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-355SS
Azide and BSA Free	NBP2-80931

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NBP2-29328

MyD88 Inhibitor Peptide Set

(3 Reviews)

(98 Publications)

2 mg

Reactivity:

Hu, Mu, Rt, Bv, Xp, Ye, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Bv, Xp, Ye, Ze

Applications:

, , , , ,

Applications: , , , , ,

Alt Catalog#

IMG-2005

Alt Catalog# IMG-2005

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NB600-936

beta Tubulin Antibody - BSA Free

(7 Reviews)

(84 Publications)

0.1 ml

NB600-936

Western Blot: beta Tubulin Antibody [NB600-936] - Analysis of extracts from HeLa, A549 and Jurkat cells using NB600-936 Beta-tubulin antibody at 1:1000

<b>Biological Strategies Validation. </b>Western Blot: beta Tubulin Antibody - BSA Free [NB600-936] - beta Tubulin Antibody [NB600-936] - Reduced levels of NADPH oxidases 1 and 4 at 18% versus 5% O2. (a) Representative Western blots showing Nox1 and Beta-tubulin in PC-3 cells or Nox4 and Beta-tubulin in C2C12 cells, at 5% and 18% O2. (b, c) Average Nox1 signal How Supraphysiological Oxygen Levels in Standard Cell Culture Affect Oxygen-Consuming Reactions. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30363917/) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: beta Tubulin Antibody [NB600-936] - PC12 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-beta Tubulin Antibody (NB600-936) 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.

Immunohistochemistry-Paraffin: beta Tubulin Antibody [NB600-936] - Analysis of FFPE tissue section of normal human skin using 1:2000 dilution of beta Tubulin antibody. Intense cytoplasmic staining of beta Tubulin (TUBB) protein was observed in various cells of the epidermal as well as the dermal cells [10X Magnification].

Western Blot: beta Tubulin Antibody [NB600-936] - Analysis in HeLa whole cell lysate at a 1:1,000 dilution.

$Western Blot: beta Tubulin Antibody [NB600-936] - Expression of pluripotency related markers and a sphere culture of BEAS-2B under normoxia. HIF-2alpha expression was detected in the nucleus of BEAS-2B under normoxia by western blotting. Beta tubulin and TATA binding protein (TBP) were used as protein markers for the cytosol and nucleus fraction, respectively. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep29311), licensed under a CC-BY license.$

Immunocytochemistry/Immunofluorescence: beta Tubulin Antibody [NB600-936] - Confocal immunofluorescent analysis of C2C12 cells using beta Tubulin antibody (NB600-936, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).

Immunocytochemistry/Immunofluorescence: beta Tubulin Antibody [NB600-936] - Analysis of beta Tubulin in mouse hippocampal primary culture. Image courtesy of product review by Lin Yi-Wen.

Immunocytochemistry/Immunofluorescence: beta Tubulin Antibody [NB600-936] - NIH3T3 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-beta Tubulin Antibody (NB600-936) 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.

Immunohistochemistry-Paraffin: beta Tubulin Antibody [NB600-936] - Analysis of FFPE tissue section of human esophageal squamous cell carcinoma (SCC) using 1:2000 dilution of beta Tubulin antibody. Strong cytoplasmic immuno-positivity of beta Tubulin (TUBB) was observed in SCC cells as well as the associated tumor stromal cells [10X Magnification].

Immunohistochemistry-Paraffin: beta Tubulin Antibody [NB600-936] - Analysis of FFPE tissue section of normal human brain using 1:2000 dilution of beta Tubulin antibody. The various brain cells depicted strong cytoplasmic immunoreactivity of beta Tubulin (TUBB) protein [10X Magnification].

Expression of AAV transgene before and after experimental autoimmune uveoretinitis (EAU) induction. (B) Western blot from retinas harvested 14 days after IRBP immunization. Membrane was probed with anti-GFP and anti-Tubulin antibodies. Image shows expression of sGFP on both sGFP and sGFP-TatM013v5 retina lysates. (n = 2-3 retina samples from different mice per group). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31795515), licensed under a CC-BY licence.

$Characterization of immunosorted NCAM1+ subpopulationA. RT-PCR analysis of gene expression in NCAM1 cell fractions. GAPDH was used as endogenous control and NCAM1− cell were used as the calibrator sample for RQ calculation (therefore = 1). Data were analysed using SDS 3.2 software and presented as average RQ ± SDEV of three replicates. ***p < 0.001, *p < 0.05 versus NCAM1−.B–D. Immunofluorescence staining of NCAM1+ and NCAM1− subpopulations for E-cadherin (E-cad) (B, red) vimentin (C, green) and SIX2 (D, green). Nuclei stained with Dapi (blue). (B–C) Images were obtained using Olympus DP72 camera attached to Olympus BX51 fluorescence microscope and processed via cellSens standard software, bar represents 200 μm. (D) Images were obtained using Zeiss LSM 510 confocal microscope, bar represents 50 μm.E. Fluorescent quantification of SIX2 immunostaining as represented in (D).F. Double labelling of sorted NCAM1+ cells for NCAM1 and SIX2: NCAM1 with DAPI (upper panel; red and blue channels), NCAM1 with SIX2 (lower panel; red and green channels), indicating both NCAM1+ SIX2+ (arrows) and NCAM1+ SIX2− (arrowheads) cells.G. Graph represents percentage of NCAM1+ SIX2+ cells and NCAM1+ SIX2− cells.H. Clonogenic efficiency of NCAM1+ cells sorted from hFK and cultured in SFM. Data are presented as average CE(%) ± SDEV. **p < 0.01 versus NCAM1−.I. Representative morphology of NCAM1+ and NCAM1− clones. Cells were observed using a Nikon Digital Sight camera attached to a Nikon Eclipse TS100 microscope.J. Clonogenic capacity of hFK cells treated with IMGN901(ADC 55 nM), huN901 (Ab55 nM) or not treated (control). Data are presented as average CE(%) ± SDEV. ***p < 0.001, *p < 0.05 versus control group. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23996934), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

PGE1 induces HIF-1 alpha protein accumulation in vascular-derived cells.Human aortic smooth muscle cells (HASMCs) (A and B), human umbilical vein endothelial cells (HUVECs) (C and D), and HEK293 cells (G) were exposed to 1 or 10 µM PGE1 under 20% O2 or 1% O2 conditions for 4 h. After treatment, cells were harvested and whole-cell lysates were subjected to immunoblot assay for HIF-1 alpha , HIF-1 beta , and beta -actin protein expression. Experiments were repeated thrice (A, C and G). Representative immunoblots are shown (A and C). Band intensities were analyzed densitometrically (B and D). Fold induction relative to lane 1 was plotted as mean ± S.D. ∗P < 0.05 compared with the control. HASMCs (E) and HUVECs (F) were exposed to 1 µM PGE1 for the indicated times under 20% O2 and were harvested for immunoblot assay for HIF-1 alpha protein. Experiments were repeated twice. Representative immunoblots are shown. (H) HASMCs and HUVECs were exposed to 10 µM PGE1 for 4 h under 20% O2 and were harvested for immunoblot assay for HIF-2 alpha protein. Experiments were repeated twice. Representative immunoblots are shown. I. HASMCs were exposed to 1 µM PGE1, lipo-PGE1 and PGE1-alfadex under 20% O2 conditions for 4 h. After treatment, cells were harvested and whole-cell lysates were subjected to immunoblot assay for HIF-1 alpha . Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24349900), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

TIP30 was regulated by HIF-2 alpha at protein level in HCC cell lines. Lentivirus infection was used to establish a stable knocking-down or overexpression of HIF-2 alpha in MHCC97H cell lines. The relationship between HIF-2 alpha and TIP30 was detected by Western blot. a Knocking-down of HIF-2 alpha upregulated TIP30 expression. b Knocking-down of TIP30 did not influence the expression of HIF-2 alpha . c, d Metformin in combination with sorafenib synergistically inhibited HIF-2 alpha protein expression and subsequently upregulated TIP30 protein expression. e MHCC97H cells were exposed to CoCl2 (400 μM) for 6 h, and anti-HIF-2 alpha or anti-IgG was used for immunoprecipitation. Immunoprecipitated and purified DNA together with 1 % of input DNA were used for PCR amplification of a 214-bp product encompassing HRE region of TIP30 promoter Image collected and cropped by CiteAb from the following open publication (//www.jhoonline.org/content/9/1/20), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

ATM and NF-kappa B activation are downregulated in Ercc1-/ delta mice heterozygous for Atm. (A) Livers were collected at 12 weeks of age from WT, Ercc1-/ delta and Ercc1-/ delta Atm+/- mic (n=3 per genotype) and lysates analyzed by western blot for activation of ATM and its downstream effectors. (B) Same liver lysates were used to measure phosphorylation of p65 and I kappa B alpha . (C) Western blot analysis of livers from 16-week-old WT, Ercc1-/ delta and Ercc1-/ delta Atm+/-mice (n=3 per genotype) probed for activation of ATM. GAPDH was used as a loading control. (D) Same liver lysates used to measure activation of NF-kappa B. (E) Fourteen-week-old livers from Ercc1-/ delta and Ercc1-/ delta p65+/- mice (n=3 per genotype) were analyzed by western blot for activation of ATM (F) and NF-kappa B. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32201398), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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

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

Reactivity:

Hu, Mu, Rt, Po, Bv, Ce, Ch, ChHa, I, In, Pm, Xp, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ce, Ch, ChHa, I, In, Pm, Xp, Ze

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-936AF350
Alexa Fluor 405	NB600-936AF405
Alexa Fluor 488	NB600-936AF488
Alexa Fluor 532	NB600-936AF532
Alexa Fluor 594	NB600-936AF594
Alexa Fluor 647	NB600-936AF647
Alexa Fluor 700	NB600-936AF700
Alexa Fluor 750	NB600-936AF750
Allophycocyanin	NB600-936APC
Biotin	NB600-936B
DyLight 350	NB600-936UV
DyLight 405	NB600-936V
DyLight 488	NB600-936G
DyLight 550	NB600-936R
DyLight 594	NB600-936DL594
DyLight 650	NB600-936C
DyLight 680	NB600-936FR
DyLight 755	NB600-936IR
FITC	NB600-936F
HRP	NB600-936H
Janelia Fluor 549	NB600-936JF549
Janelia Fluor 646	NB600-936JF646
PE	NB600-936PE
PerCP	NB600-936PCP
mFluor Violet 450 SE	NB600-936MFV450
mFluor Violet 500 SE	NB600-936MFV500
mFluor Violet 610 SE	NB600-936MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-936SS

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NBP1-30054

Collagen I alpha 1 Antibody - Azide and BSA Free

(2 Reviews)

(67 Publications)

0.1 ml

NBP1-30054

Immunohistochemistry: Collagen I alpha 1 Antibody - Azide and BSA Free [NBP1-30054] - Collagen I Antibody [NBP1-30054] - Formaldehyde-fixed fibrotic mouse lung tissue. The antibody recognizes mature collagen I (red) that has formed fibrils in the extracellular matrix.

Simple Western: Collagen I alpha 1 Antibody - Azide and BSA Free [NBP1-30054] - Simple Western lane view shows a specific band for Collagen I Alpha 1 in 0.5 mg/ml of Human Lung (left) and Human Kidney (right) lysate. This experiment was performed under reducing conditions using the 66-440 kDa separation system. * Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody

TNC deficiency reduced kidney fibrosis in animal models. D) Western blot showed that the proteins of collagen Ialpha and alpha-SMA were also significantly reduced in TNC-/- mice at UUO day 7 (C, n = 7, p < 0.05). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/36522320), licensed under a CC-BY licence.

Immunohistochemistry staining for type I collagen. Immunohistochemical staining targeting type I collagen revealed severe disorganization of the annulus fibrosus (AF), and a decrease in staining intensity for group D. On the contrary, the arc-shaped and layered AF structure with immunoreactivity for type I collagen staining were mostly maintained in group S when compared to group C. Presented images show the AF at site of surgery, defect, and transplant. Abbreviations: D: AF defect only group, S: AF defect treated with sheet transplantation group, C: sham group, NC: negative control of sample stained without primary antibody (scale bar = 300 um) Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31463464), licensed under a CC-BY licence.

Reactivity:

Hu, Mu, Rt, Am, Av, Ma, Sh, Bv, Ca, Ch, Eq, Fe, Av-Bi, Gt, Gp, Ha, Pm, Rb, Xp

Species Glossary

Reactivity: Hu, Mu, Rt, Am, Av, Ma, Sh, Bv, Ca, Ch, Eq, Fe, Av-Bi, Gt, Gp, Ha, Pm, Rb, Xp

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Azide and BSA Free	NBP1-30054-0.02ml

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NB600-532

beta-Actin Antibody

(6 Reviews)

(63 Publications)

0.1 mg

NB600-532

Western Blot: beta-Actin Antibody [NB600-532] - Detection of human and mouse Cytoskeletal Actin by western blot. Samples: Whole cell lysate (50 ug) from HeLa, HEK293T, Jurkat, mouse TCMK-1, and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-Cytoskeletal Actin antibody NB600-532 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 1 second.

Western Blot: beta-Actin Antibody [NB600-532] - Mouse colon whole cell lysate. PVDF membrane was probed with Rabbit Anti-B actin Antibody (Catalog # NB600-532) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody. WB image submitted by a verified customer review.

Immunohistochemistry: beta-Actin Antibody [NB600-532] - Detection of human Cytoskeletal Actin by immunohistochemistry. Sample: FFPE section of human ovarian cancer. Antibody: Affinity purified rabbit anti-Cytoskeletal Actin (NB600-532). Detection: DAB

Western Blot: beta-Actin Antibody [NB600-532] - Western blot analysis of Actin (NB600-532) using RCC4 whole cell lysate [NBP1-30412].

Western Blot: beta-Actin Antibody [NB600-532] - Western blot analysis of Actin (NB600-532) using HepG2 whole cell lysate [NBP1-42569].

Western Blot: beta-Actin Antibody [NB600-532] - Analysis using the HRP conjugate of NB600-532. Detection of Beta Actin in MCF-7 cell lysate (20ug) using anti-Beta Actin antibody. WB image submitted by a verified customer review.

Western Blot: beta-Actin Antibody [NB600-532] - Treatments of periplocin and/or TRAIL activate DR4, FADD, and proapoptotic proteins in HCC cells. The effect of periplocin treatment on the expression of DR4, DR5, and FADD was analyzed by western blot. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/ecam/2013/958025/), licensed under a CC-BY license.

Western Blot: beta-Actin Antibody [NB600-532] - Hypoxic preconditioning increases PBMNC sensitivity to apelin-13 via upregulation of APJ expression, leading to growth factor secretion.(a) APJ protein expression was significantly increased in hypoxic PBMNCs. *p <0.05 vs. N-PBMNCs. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep19379), licensed under a CC-BY license.

Immunohistochemistry: beta-Actin Antibody [NB600-532] - Sample: FFPE section of human lung carcinoma. Antibody: Affinity purified rabbit anti-Cytoskeletal Actin used at a dilution of 1:1,000 (1ug/ml). Detection: DAB

Hypoxic preconditioning increases PBMNC sensitivity to apelin-13 via upregulation of APJ expression, leading to growth factor secretion.(a) APJ protein expression was significantly increased in hypoxic PBMNCs. *p < 0.05 vs. N-PBMNCs. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26763337), licensed under a CC-BY licence.

L. donovani infection induces HIF-1 alpha expression in CD11chi splenic DCs in an IRF-5 dependent manner.Mice were infected with 2x107 amastigotes intravenously. (A) Real-time PCR analysis of HIF-1 alpha mRNA expression in CD11c+ cells purified from C57BL/6 mice at various time points after infection. (B) Immunoblot analysis of HIF-1 alpha expression in CD11c+ cells from C57BL/6 mice (upper panel) and densitometric analysis normalized to beta -actin expression and expressed as fold increase to results obtained with naïve mice (lower panel). (C) Real-time PCR analysis of HIF-1 alpha expression in sorted CD11c+ cells from Irf-5flox/floxCre- and Irf-5flox/floxCMV-Cre+. (D) Immunoblot analysis of Hif-1 alpha expression in CD11c+ cells population of Irf5flox/floxCre- (left upper panel) and Irf-5flox/floxCMV-Cre+ (right upper panel), and densitometric analysis normalized to beta -actin expression and expressed as fold increase to results obtained with naïve mice (lower panels). (E) Real-time PCR analysis of Hif-1 alpha expression in CD11c- splenocytes from Irf-5flox/floxCre- and Irf-5flox/floxCMV-Cre+. (F) Real-time PCR analysis of HIF-1 alpha mRNA expression in BMDC from Irf-5flox/floxCMV-Cre+ and Cre- mice. All data represent mean ± SEM combined from 3 independent experiments. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.ppat.1004938), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Effects of autophagy inhibitors in combination with IFN-gamma on expression of PD-L1 in gastric cancer cell lines. a The effect of chloroquine (CQ) or 3-MA on expression of PD-L1 with or without INF-gamma for 24 h was determined by flow cytometry assays. In AGS and NCI-n87 cells, MFI as the indication of PD-L1 expression level can be further increased by the treatment of INF-gamma . b Levels of LC3B-I/II, p62/SQSTM1 and PD-L1 were determined by Western blots in AGS and NCI-n87 cells treated by CQ, 3-MA, bafilomycin A1 (Baf) or rapamycin (Rap) for 24 h. c Positive staining of PD-L1 (red) and LC3 positive puncta (green) was determined by immunofluorescence in AGS and NCI-n87 cells treated by autophagy inhibitors and activator as in (b). d Rapamycin decreased the levels of PD-L1 protein in AGS and NCI-n87 cells as shown by flow cytometry. Results were averaged and blots were representative of 4 independent experiments, *p < 0.05, **p < 0.01 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30925913), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ca, Ch, Pm, SyHa, Gp, Pm-Or, Rb, Sh, Xp

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Ch, Pm, SyHa, Gp, Pm-Or, Rb, Sh, Xp

Applications:

, , , ,

Applications: , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-532AF350
Alexa Fluor 405	NB600-532AF405
Alexa Fluor 488	NB600-532AF488
Alexa Fluor 532	NB600-532AF532
Alexa Fluor 594	NB600-532AF594
Alexa Fluor 647	NB600-532AF647
Alexa Fluor 700	NB600-532AF700
Alexa Fluor 750	NB600-532AF750
Biotin	NB600-532B
DyLight 350	NB600-532UV
DyLight 405	NB600-532V
DyLight 488	NB600-532G
DyLight 550	NB600-532R
DyLight 594	NB600-532DL594
DyLight 650	NB600-532C
DyLight 680	NB600-532FR
DyLight 755	NB600-532IR
FITC	NB600-532F
HRP	NB600-532H
Janelia Fluor 549	NB600-532JF549
Janelia Fluor 646	NB600-532JF646

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NB100-1965

Calnexin Antibody - BSA Free

(3 Reviews)

(39 Publications)

0.1 ml

NB100-1965

$Western Blot: Calnexin Antibody [NB100-1965] - Subcellular localization of TM6SF2.Immunoaffinity isolation of ER and Golgi complex from mouse liver. ER and Golgi fractions were prepared from mouse liver microsomes by immunoaffinity chromatography as described under Experimental Procedures. Microsome membranes were dissolved in RIPA buffer, and equal volumes were separated on 10% SDS-PAGE and immunoblotting as described under Experimental Procedures. BiP, binding immunoglobulin protein; Gos28, Golgi SNAP receptor complex member 1; *, nonspecific band. Image collected and cropped by Citeab from the following publication (Inactivation of Tm6sf2, a Gene Defective in Fatty Liver Disease, Impairs Lipidation but Not Secretion of Very Low Density Lipoproteins. J Biol Chem (2016) licensed under a CC-BY license.$

Immunohistochemistry: Calnexin Antibody [NB100-1965] - Calnexin immunostaining in the anterior (AI) and mid (MI) intestine of ctrl (control zebrafish), D.I.O. (diet-induced obesity zebrafish), D.I.O. flw 3,5-T2 (D.I.O. zebrafish followed by 3,5-T2), D.I.O. with 3,5-T2 (D.I.O. zebrafish treated with 3,5-T2). (A) AI of ctrl zebrafish (B) AI of D.I.O. (C) AI of D.I.O. flw 3,5-T2. (D) AI of D.I.O. with 3,5-T2. The arrows indicate calnexin immunoexpression in the enteroendocrine and goblet cells. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2076-2615/10/7/1131/htm#) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: Calnexin Antibody [NB100-1965] - HeLa 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 (NB100-1965) at 1 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.

Immunocytochemistry/Immunofluorescence: Calnexin Antibody [NB100-1965] - NIH3T3 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 (NB100-1965) at 1 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.

Immunocytochemistry/Immunofluorescence: Calnexin Antibody [NB100-1965] - Rat FR 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 Calnexin Antibody (NB100-1965) at 1 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.

Western Blot: Calnexin Antibody - BSA Free [NB100-1965] - Whole cell lysate or exosome from MDA-MD-231 cells was loaded with 20 ug/lane. 10% SDS-PAGE. Calnexin Antibody (NB100-1965) was used for primary antibody: 1:3000, 4C, overnight. Image from verified customer review.

Flow Cytometry: Calnexin Antibody [NB100-1965] - An intracellular stain was performed on rat FR cells with Calnexin Antibody NB100-1965 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Western Blot: Calnexin Antibody [NB100-1965] - A dilution of 1/2000 used in TBS with 0.05% Tween-20. Bands visualized by ECL using goat anti-rabbit HRP at 1:2000.

Immunohistochemistry: Calnexin Antibody [NB100-1965] - IHC analysis of CANX in mouse prostate using DAB with hematoxylin counterstain.

Flow Cytometry: Calnexin Antibody [NB100-1965] - An intracellular stain was performed on Daudi cells with Calnexin Antibody NB100-1965 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Flow Cytometry: Calnexin Antibody [NB100-1965] - An intracellular stain was performed on NIH3T3 cells with Calnexin Antibody NB100-1965 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Simple Western: Calnexin Antibody [NB100-1965] - Simple Western lane view shows a specific band for Calnexin in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Calnexin immunostaining in the anterior (AI) and mid (MI) intestine of ctrl (control zebrafish), D.I.O. (diet-induced obesity zebrafish), D.I.O. flw 3,5-T2 (D.I.O. zebrafish followed by 3,5-T2), D.I.O. with 3,5-T2 (D.I.O. zebrafish treated with 3,5-T2). (A) AI of ctrl zebrafish (B) AI of D.I.O. (C) AI of D.I.O. flw 3,5-T2. (D) AI of D.I.O. with 3,5-T2. (E) MI of ctrl zebrafish. (F) MI of D.I.O. (G) MI of D.I.O. flw 3,5-T2. and (H) MI of D.I.O. with 3,5-T2. The arrows indicate calnexin immunoexpression in the enteroendocrine and goblet cells. (I,J) Bar graphs showing calnexin optical density (O.D.) in the (I) AI and (J) MI of ctrl, D.I.O., D.I.O. flw 3,5-T2 and D.I.O. with 3,5-T2. Data are expressed as mean +/- SE. ** p < 0.001, * p < 0.05 compared to the control group. p < 0.05 compared to D.I.O. Scale bar: 100 um in the low magnification and 50 um in the higher magnification in the boxes. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32635261), licensed under a CC-BY licence.

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

Reactivity:

Hu, Mu, Rt, Po, Av, Bv, Ch, Dr, Gp, Rb, Sh, Xp, Ze, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Av, Bv, Ch, Dr, Gp, Rb, Sh, Xp, Ze, Ca

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	NB100-1965AF350
Alexa Fluor 405	NB100-1965AF405
Alexa Fluor 488	NB100-1965AF488
Alexa Fluor 532	NB100-1965AF532
Alexa Fluor 594	NB100-1965AF594
Alexa Fluor 647	NB100-1965AF647
Alexa Fluor 700	NB100-1965AF700
Alexa Fluor 750	NB100-1965AF750
Allophycocyanin	NB100-1965APC
Biotin	NB100-1965B
DyLight 350	NB100-1965UV
DyLight 405	NB100-1965V
DyLight 488	NB100-1965G
DyLight 550	NB100-1965R
DyLight 594	NB100-1965DL594
DyLight 650	NB100-1965C
DyLight 680	NB100-1965FR
DyLight 755	NB100-1965IR
FITC	NB100-1965F
HRP	NB100-1965H
Janelia Fluor 549	NB100-1965JF549
Janelia Fluor 646	NB100-1965JF646
PE	NB100-1965PE
PerCP	NB100-1965PCP
mFluor Violet 450 SE	NB100-1965MFV450
mFluor Violet 500 SE	NB100-1965MFV500
mFluor Violet 610 SE	NB100-1965MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-1965SS

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

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

Marker

Modification

Clonality

Clone

Gene Symbol

Images Type

Inhibitor Family

Inhibitor Target

Isotype

Preservative

Research Areas

Has Sample Size

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Species Glossary

HIF-1 alpha Antibody (H1alpha67)

LC3B Antibody (1251A) - BSA Free

ATG5 Antibody - BSA Free

HIF-1 alpha Antibody - BSA Free

alpha Tubulin Antibody (DM1A) - BSA Free

Sodium Potassium ATPase Alpha 1 Antibody (464.6)

RPE65 Antibody (401.8B11.3D9) - BSA Free

MyD88 Inhibitor Peptide Set

beta Tubulin Antibody - BSA Free

Collagen I alpha 1 Antibody - Azide and BSA Free

beta-Actin Antibody

Calnexin Antibody - BSA Free