Signal Transduction reagents: Novus Biologicals

NB400-104

SR-BI Antibody - BSA Free

(5 Reviews)

(243 Publications)

0.1 ml

NB400-104

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-104] - SR-BI antibody was tested in human fibroblast samples fixed in 4% PFA and permeabilized in PBS (0.2% Tween). Primary incubation overnight at 4C using a 1:100 dilution in PBS (0.1% Tween) with 1% BSA. Secondary antibody is anti-rabbit conjugate to Alexa Fluor 488. SR-BI is shown in green and nucleus in blue (Hoescht 33342 stain).

Flow Cytometry: SR-BI Antibody [NB400-104] - An intracellular stain was performed on HeLa cells with SR-BI antibody NB400-104AF488 (blue) and a matched isotype control NBP2-24893AF488 (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.

Western Blot: SR-BI Antibody [NB400-104] - Detection of SR-BI in rat H4IIE total cell lysates and plasma membrane proteins. Photo courtesy of product review by verified customer.

Immunohistochemistry: SR-BI Antibody [NB400-104] - Immunolocalization of SR-BI in adult mink testis using NB400-104. SR-BI labeling is visible at the surface and along the outline of the large vacuole. Photo courtesy of R.M. Pelletier, University of Montreal.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - FFPE tissue section of mouse liver using SR-BI antibody (Lot 8310) at 1:300 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a specific membrane signal of SR-BI protein in the murine hepatocytes.

<b>Genetic Strategies Validation. </b>Western Blot: SR-BI Antibody [NB400-104] - HCV replication was increased by CD36. Silence of CD36 decreased HCV infection and replication (n = 3). Huh7.5 cells were transfected with 0.2 u g CD36-HA plasmid or with 150 pmol siRNA for CD36 (B) for 48 hrs, and then were washed and infected with HCV (150 IU/cell) for 2 hrs, followed by washing and continuously incubating. Intracellular HCV RNA and proteins were detected in 96 hrs, and the cytotoxicity of siRNAs was measured in 96-wells plate with a MTT assay in B. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21808) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - Staining of human adrenal cortex.

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-104] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-SR-BI at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - FFPE tissue section of mouse liver using SR-BI antibody (Lot R-4) at 1:300 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated mainly a membranous signal of SR-BI protein in the murine hepatocytes.

Flow Cytometry: SR-BI Antibody [NB400-104] - Analysis of Huh7 and HepG2 cells using SR-B1 antibody NB400-104. Courtesy of Bruno Sainz, Jr., PhD, University of Illinois at Chicago.

ABCA1, ABCG1 & SR-BI protein expressions. a & b Hepatic protein expressions of ABCA1, ABCG1 & SR-BI were significantly decreased in COMT−/− mice at GD 18.5, compared to C57BL/6 J mice. Decreased hepatic ABCA1 expression was also observed at 10 days postpartum. ATI-5261 increased ABCA1 & ABCG1 expression in the liver at 10 days postpartum. c Placental protein expressions of ABCA1 & ABCG1 was reduced in COMT−/− mice, compared to C57BL/6 J mice. ATI-5261 treatment significantly increased ABCA1 levels in the placenta of COMT−/− mice. d Representative immunoblots of the corresponding proteins in the placenta with mouse RAW264.7 cell lysate included as positive control. Similar results were obtained when the experiment was repeated with lysates prepared from three batches of tissues. Data are presented as mean ± SEM. Groups (n = 8 in all groups) were compared using one-way ANOVA with post-hoc analysis (Tukey’s procedure). *, p 2000) (g, h, i, respectively). Bar graph shows the mean percentage of positively stained cells. Error bars depict standard deviations. ****p 0.3) by one-way ANOVA. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29523748), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SR-BI Antibody - BSA Free [NB400-104] - Effects of 13-HODE & LA in the presence & absence of PPAR alpha & PPAR gamma selective antagonists on molecular markers of cholesterol homeostasis in RAW264.7 macrophages. RAW264.7 cells were pre-treated without or with the PPAR alpha selective antagonist GW6471 or the PPAR gamma selective antagonist GW9662 & subsequently treated without (vehicle control) or with 2.5 μmol/L 13-HODE or 100 μmol/L LA for 24 h. Afterwards, cells were lysed & subsequently processed for western blotting as described in the materials & methods section. A, Representative immunoblots specific for ABCA1, ABCG1, SR-BI, LXR alpha , & beta -actin which was used for normalization are shown. B, Bars represent data from densitometric analysis & are means ± SD from three independent experiments (n = 3). Data are expressed as percentage of protein concentration of vehicle control cells. Results from statistical analysis are indicated: Significant effects are denoted with superscript letters. Bars marked without a common superscript letter significantly differ (P 2000) (g, h, i, respectively). Bar graph shows the mean percentage of positively stained cells. Error bars depict standard deviations. ****p 0.3) by one-way ANOVA. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29523748), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SR-BI Antibody - BSA Free [NB400-104] - Differentiation & characteristics of iHLCs. (a) Cell morphology & culture conditions at different stages of differentiation from iPSCs into iHLCs. Bright-field images were taken at the indicated time points (scale bar 100 µm). (b) Sequential expression & repression of transcription factors marking successful differentiation. Cells were immunostained with antibodies against Oct 3/4 (pluripotency marker), the endodermal transcription factor GATA-4, & the hepatic marker HNF4 alpha at day 0, 5, & 10 of differentiation (scale bars 20 µm). Shown are representative fluorescence & DIC images. (c) mRNA expression levels of different hepatocyte-specific factors in iHLCs, Huh7.5 cells, & primary human hepatocytes (PHHs) were analysed via qRT-PCR. Expression levels are displayed as fold over iPSCs normalised to GAPDH & 18S rRNA (mean ± SEM, n = 3–4, *p 0.9) by one-way ANOVA. §P < 0.02 by unpaired Student's t-test, relative to 0 h time point. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29523748), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, ChHa, Ha, Md, Pm, Rb, SyHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, ChHa, Ha, Md, Pm, Rb, SyHa

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-104AF350
Alexa Fluor 405	NB400-104AF405
Alexa Fluor 488	NB400-104AF488
Alexa Fluor 532	NB400-104AF532
Alexa Fluor 594	NB400-104AF594
Alexa Fluor 647	NB400-104AF647
Alexa Fluor 700	NB400-104AF700
Alexa Fluor 750	NB400-104AF750
Allophycocyanin	NB400-104APC
Biotin	NB400-104B
CoraFluor 1	NB400-104CL1
DyLight 350	NB400-104UV
DyLight 405	NB400-104V
DyLight 488	NB400-104G
DyLight 550	NB400-104R
DyLight 594	NB400-104DL594
DyLight 650	NB400-104C
DyLight 680	NB400-104FR
DyLight 755	NB400-104IR
FITC	NB400-104F
HRP	NB400-104H
Janelia Fluor 525	NB400-104JF525
Janelia Fluor 549	NB400-104JF549
Janelia Fluor 646	NB400-104JF646
Janelia Fluor 669	NB400-104JF669
PE	NB400-104PE
PerCP	NB400-104PCP
mFluor Violet 450 SE	NB400-104MFV450
mFluor Violet 500 SE	NB400-104MFV500
mFluor Violet 610 SE	NB400-104MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-104SS

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NB100-134

HIF-1 alpha Antibody - BSA Free

(14 Reviews)

(236 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).

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 & are expressed relative to control (n = 3). (B,C) Increased Wnt11 mRNA by hypoxia in EMSC preadipocytes & adipocytes (Day 0–12 after differentiation) (B), & C2C12 myoblast & myocyte (Day 0 & 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 & 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) & 4T1 cells (F). (G) EMSCs treated with 0.1 mM DMOG for the indicated times. Wnt11 & Vegf mRNA expression was measured by qPCR & 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 25 pictures/group from three separate experiments. Scale bar = 40 µm. (F) Cell proliferation was assessed by evaluating PCNA expression, n = 4. * p 20). B May–Grunwald–Giemsa staining of whole KM blood cells in 6-month-old AB zebrafish after 4 days of E2 treatment (t-test, ***p 20). F Effect of E2 on neutrophil proliferation in AB zebrafish embryos. (one-way ANOVA (LSD). ns, no significance, n > 10). G E2 promotes the apoptosis of myeloid lineage cells in AB zebrafish embryos (one-way ANOVA (LSD) **p 10). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35842445), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - IGFBP5 inhibits HIF1 alpha expression through the MAPK-ERK signaling pathwayA. & B. Western blots using specific antibodies for the phosphorylation state of IGF1R, ERK1/2, & p38-MAPK from A375 IGFBP5 OE & control cells. Phosphorylation of IGF1R, ERK1/2, & p38-MAPK was decreased in A375 cells transfected with IGFBP5. GAPDH was used as a loading control. C. Overexpression of IGFBP5 inhibited HIF1 alpha expression visualized by western blots of A375 cells. The arrow points to the band of HIF1 alpha . D. Assessment of the reduced gene expression levels of VEGFA & MMP9, downstream genes regulated by HIF1 alpha , 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 & vector control cells. *, P < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26010068), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - WNT11 regulates MMPs activities.(A–C) (Top panels): Serum-free medium was conditioned for 24 hrs by the indicated cells, concentrated 20-fold & assayed by gelatin zymography. Gelatinolytic activity is indicated by clear zones against a dark background of stained substrate. (Bottom): Whole cell extracts were immunoblotted with indicated antibodies. (A) Overexpression of Wnt11 in EMSC or BT473 cells enhances activity of MMP-9 & MMP-2. (B) Impaired activity of MMP-9 & MMP-2 in MDA-MB-231 cells (left) or EMSCs (right) stably expressing Wnt11 shRNAs & treated with DMOG. (C) WNT11 is required for MMP-9 & MMP-2 activity in MDA-MB-231 cells (left) or EMSCs (right) under normoxic & hypoxic culture conditions. (D) WNT11 regulates MMP2 protein in media. (Top): conditioned media from indicated cells & treatments. (Bottom): whole cell lysates were immunoblotted with indicated antibodies. (E) Recombinant WNT11 induces both MMP-2 protein & MMP-2 activity in media. (Top panels): Gelatin zymography & immunoblot of serum-free medium conditioned for the indicated times after recombinant WNT11 (r-WNT11) treatment. (Bottom): Whole cell lysates were immunoblotted with indicated antibodies. (F) MMP-2 inhibitor attenuated induced migration by WNT11. MDA-MB-231 cells infected with lentiviruses for stable expression of Wnt11 or GFP (n = 4) were incubated with either vehicle or 1 μM of ARP100. Media in the lower compartment had same concentration of DMSO or inhibitor. Values are mean ± s.e.m. *p < 0.05, **p < 0.01. For panels (A–D), HIF-1 alpha & HIF-2 alpha were shown as a marker of hypoxia, WNT11 normalized to alpha -Tubulin was shown. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - ER-INP binds to PHD2, inhibits hydroxylation of HIF-1 alpha & increases HIF-1 alpha accumulation. (A) Co-immunoprecipitation assay. HEK293 cells were transfected with control vector or intrabody ER-INP for 48 hours & subsequently lysed. Co-immunoprecipitation & western blot assays were performed on the cell lysis. ER-INP recognized & bound to PHD2 in HEK293 cells (n = 3). (B) Western blot analysis to measure the effect of ER-INP on HIF-1 alpha & its hydroxylation level in transfected HEK293 & RAW264.7 cells pre-treated with MG132 (upper panel) & the protein ratio to beta -actin loading control by ImageJ densitometry analysis (lower panel; n = 3). (C) Immunofluorescence assay. Expression of ER-INP increases HIF-1 alpha accumulation in RAW264.7 cells. Cells were stained with anti-HIF-1 alpha antibody & DAPI & then visualized & photographed under immunofluorescence microscopy (left), & mean fluorescence intensity of HIF-1 alpha versus the mean fluorescence intensity of nuclear DAPI staining is displayed (right). Data represent the mean ± SD of 3 independent slides. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31413262), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - HIF-1 alpha binds to TIM-3 promoter & regulates its expression in primary glia.(a) Cell surface expression of TIM-3 analysed in BV2 cells under 20% O2 or 1% O2 for 24 h by flow cytometry using PE-conjugated anti-TIM-3 antibody. Results from 3 independent experiments presented as a representative histogram & the mean fold change (± s.d.) relative to normoxic sample. (b) Mouse primary mixed glial cells incubated under hypoxia or normoxia for 24 h, & the cells examined by immunocytochemistry using an anti-TIM-3 antibody. (c,d) Mouse primary mixed glial cells & primary neuronal cells incubated under hypoxia or normoxia for 24 h, & then RT–PCR used to detect the levels of TIM-3 & actin. Relative transcript levels shown as the mean fold change (± s.d.) from 3 independent experiments (NS, not significant, Student–Newman–Keuls test). (e) Primary mixed glial cells incubated under hypoxia or normoxia for 24 h, & chromatin immunoprecipitation (ChIP) performed w/ anti-HIF-1 alpha or control IgG. Results presented as relative amounts representative of 3 independent experiments. (f) Primary mixed glial cells cultured from HIF-1 alpha +f/+f mice, infected w/ Ad-GFP or Ad-Cre/GFP, transfected w/ TIM-3-luciferase reporter constructs & incubated under hypoxic or normoxic conditions for 24 h. Relative promoter activity is expressed as the ratio of luciferase activity/ beta -galactosidase activity. (g,h) RT–PCR (g) & WB analysis (h) performed under hypoxia or normoxia for 24 h using the indicated primers & antibodies, respectively. The data shown representative of at least 3 independent experiments. The graphs show the % changes in TIM-3 transcript & protein levels in Ad-Cre/GFP- versus Ad-GFP-infected cells under hypoxia. IP, immunoprecipitation. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25790768), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - Regulation of catabolic genes by HIF-1 alpha . (A) Protein levels of Hif1a, & mRNA levels of Mmp13, & Hif2a in WT mouse primary chondrocytes transduced with GFP or HIF1A adenoviral vectors under the hypoxic condition (3% O2). GFP or HIF1A was transduced at a multiplicity of infection (MOI) of 100. The cells were treated with or without 10 ng/mL IL-1 beta for 2 days. Bars show the mean ± SD of three samples per group. *P < 0.05. (B) Protein levels of Hif1a, & mRNA levels of Mmp13, & Hif2a in WT mouse primary chondrocytes transfected with siRNA against GFP or Hif1a under the hypoxic condition. The cells were treated with or without 10 ng/mL IL-1 beta for 2 days. Bars show the mean ± SD of three samples per group. *P < 0.05. (C) Protein levels of Hif1a, & mRNA levels of Mmp13, & Hif2a in Hif1afl/fl primary chondrocytes transduced with GFP or Cre adenoviral vectors under the hypoxic condition. GFP or Cre was transduced at a MOI of 100. The cells were treated with or without 10 ng/mL IL-1 beta for 2 days. Bars show the mean ± SD of three samples per group. *P < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32214220), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - NSCs Reduce Succinate Levels & Reprogram the Metabolism of Type 1 Pro-inflammatory Mφ toward Oxidative Phosphorylation In Vitro(A) Experimental setup for in vitro MφLPS co-cultures with iNSCs/NSCs.(B & C) Gene expression microarrays of MφLPS-iNSCs/NSCs. (B) Venn diagram of differentially expressed genes (adjusted p value < 0.1). (C) Heatmap of genes differentially expressed (adjusted p value < 0.1) in MφLPS-iNSCs or MφLPS-NSCs.(D & E) qRT-PCR independent validation of differentially expressed inflammatory genes as in (C). (D) Expression of genes related to type 1 inflammatory (E) & anti-inflammatory Mφ phenotypes relative to Actb. Data are mean fold change (±SEM) versus MφLPS from n ≥ 3 independent replicates per condition.(F) qRT-PCR of BV2LPS-iNSCs/NSCs (±SEM) from n ≥ 3 independent experiments per condition. BV2 & BV2LPS are shown as controls.(G & H) Extracellular flux (XF) assay of the oxygen consumption rate (OCR) (G) & extracellular acidification rate (ECAR) (H) in MφLPS-iNSCs/NSCs. Data were normalized on total protein content & are expressed as mean values (±SEM) from n ≥ 3 independent experiments per condition.(I & J) Levels of significantly changed extracellular (EXTRA_Metab, I) & intracellular (INTRA_Metab, J) metabolites in MφLPS versus Mφ at 25 hr. Data are mean a.u. (±SEM) from n ≥ 2 independent experiments per condition.(K & L) Hif-1 alpha (K), PKM2 (K), & IL-1 beta (L) expression levels relative to beta -actin. Data are mean fold change versus MφLPS (±SEM) from n ≥ 3 independent experiments per condition.∗p ≤ 0.05 & ∗∗p ≤ 0.01 versus MφLPS. See also Tables S2 & S3. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29478844), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - Enhanced expression of hypoxia-induced survival factors & angiogenic growth factors in hASC L-spheroids.(A) The light source used was LED (660 nm) designed to fit over a microplate (12.5 × 8.5 cm) for cell culture. (B) Formation of hASC L-spheroids. hASCs morphology on non–tissue culture–treated 24-well plates at day 3. Scale bar = 500 μm. (C) Western blot analysis & quantification of HIF1-alpha in hASCs cultured as spheroids, L-spheroids & monolayers (*p < 0.01, compared to the L-spheroid group). (D) Angiogenesis-related protein analysis of L-spheroids (*, p < 0.05, compared to the spheroid group, t-test, n = 3 in each group). (E) ELISA measurement of spheroids cultured for 3 days. Concentrations of VEGF are presented as pg-corrected for 104 cells. (*, p < 0.05, compared with spheroid 6J/cm2 group, t-test, n = 3 in each group). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26065900), 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
CoraFluor 1	NB100-134CL1
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 525	NB100-134JF525
Janelia Fluor 549	NB100-134JF549
Janelia Fluor 646	NB100-134JF646
Janelia Fluor 669	NB100-134JF669
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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(select up to 3 products from this page to compare)

NBP2-75977

HIF-1 alpha Antibody (2443B) - BSA Free

0.1 mg

NBP2-75977

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (2443B) [NBP2-75977] - Immunofluorescent staining of HIF-1 alpha using catalog number NBP2-75977 (clone 2443B). Staining was observed in Hela cells treated with DFO but not in untreated Hela cells.

Simple Western: HIF-1 alpha Antibody (2443B) [NBP2-75977] - Lane view shows a specific band for HIF-1 alpha in HepG2 +/- DFO cell lysate using 20ug/ml. Electropherogram image of corresponding Simple Western lane view at WES molecular weight of 116 KDa. Image from an internal validation.

Simple Western: HIF-1 alpha Antibody (2443B) [NBP2-75977] - 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 NBP2-75977 (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.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

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	NBP2-75977AF350
Alexa Fluor 405	NBP2-75977AF405
Alexa Fluor 488	NBP2-75977AF488
Alexa Fluor 532	NBP2-75977AF532
Alexa Fluor 594	NBP2-75977AF594
Alexa Fluor 647	NBP2-75977AF647
Alexa Fluor 700	NBP2-75977AF700
Alexa Fluor 750	NBP2-75977AF750
Allophycocyanin	NBP2-75977APC
Biotin	NBP2-75977B
CoraFluor 1	NBP2-75977CL1
DyLight 350	NBP2-75977UV
DyLight 405	NBP2-75977V
DyLight 488	NBP2-75977G
DyLight 550	NBP2-75977R
DyLight 594	NBP2-75977DL594
DyLight 650	NBP2-75977C
DyLight 680	NBP2-75977FR
DyLight 755	NBP2-75977IR
FITC	NBP2-75977F
HRP	NBP2-75977H
Janelia Fluor 525	NBP2-75977JF525
Janelia Fluor 549	NBP2-75977JF549
Janelia Fluor 646	NBP2-75977JF646
Janelia Fluor 669	NBP2-75977JF669
PE	NBP2-75977PE
PerCP	NBP2-75977PCP
mFluor Violet 450 SE	NBP2-75977MFV450
mFluor Violet 500 SE	NBP2-75977MFV500
mFluor Violet 610 SE	NBP2-75977MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-75977SS
Azide and BSA Free	NBP2-80764

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NBP2-75978

HIF-1 alpha Antibody (2443C) - BSA Free

(2 Reviews)

(1 Publication)

0.1 mg

NBP2-75978

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (2443C) [NBP2-75978] - Immunofluorescent staining of HIF-1 alpha using catalog number NBP2-75977 (clone 2443B). Staining was observed in Hela cells treated with DFO but not in untreated Hela cells.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (2443C) [NBP2-75978] - HIF-1 alpha (clone 2443C) was detected in immersion fixed paraffin-embedded sections of human kidney using Rabbit Anti-Human HIF-1 alpha Monoclonal Antibody (Catalog # NBP2-75978) at 1.0 ug/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte (TM) HRP Polymer Antibody (Catalog # VC002). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nuclei.

Simple Western: HIF-1 alpha Antibody (2443C) [NBP2-75978] - Lane view shows a specific band for HIF-1 alpha in HepG2 +/- DFO cell lysate using 20 ug/mL. Electropherogram image of corresponding Simple Western lane view at WES molecular weight of 116 KDa. Image from an internal validation.

Simple Western: HIF-1 alpha Antibody (2443C) [NBP2-75978] - 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 NBP2-75978 (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.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

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	NBP2-75978AF350
Alexa Fluor 405	NBP2-75978AF405
Alexa Fluor 488	NBP2-75978AF488
Alexa Fluor 532	NBP2-75978AF532
Alexa Fluor 594	NBP2-75978AF594
Alexa Fluor 647	NBP2-75978AF647
Alexa Fluor 700	NBP2-75978AF700
Alexa Fluor 750	NBP2-75978AF750
Allophycocyanin	NBP2-75978APC
Biotin	NBP2-75978B
CoraFluor 1	NBP2-75978CL1
DyLight 350	NBP2-75978UV
DyLight 405	NBP2-75978V
DyLight 488	NBP2-75978G
DyLight 550	NBP2-75978R
DyLight 594	NBP2-75978DL594
DyLight 650	NBP2-75978C
DyLight 680	NBP2-75978FR
DyLight 755	NBP2-75978IR
FITC	NBP2-75978F
HRP	NBP2-75978H
Janelia Fluor 525	NBP2-75978JF525
Janelia Fluor 549	NBP2-75978JF549
Janelia Fluor 646	NBP2-75978JF646
Janelia Fluor 669	NBP2-75978JF669
PE	NBP2-75978PE
PerCP	NBP2-75978PCP
mFluor Violet 450 SE	NBP2-75978MFV450
mFluor Violet 500 SE	NBP2-75978MFV500
mFluor Violet 610 SE	NBP2-75978MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-75978SS
Azide and BSA Free	NBP2-80765

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NB100-56708

Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free

(2 Reviews)

(202 Publications)

0.1 mg

NB100-56708

Image of Caspase-3 Antibody (31A1067) - (Pro and Active). Whole cell protein from Jurkat cells treated with and without 2 uM staurosporine as indicated was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 5 ug/ml anti-Caspase 3 in 1% milk, and detected with an anti-mouse HRP secondary antibody using a Femto sensitivity chemiluminescence reagent. Note the detection of both pro-caspase 3 at 35 kDa and the cleaved active caspase 3 at 15-17 kDa.

<b>Biological Strategies Validation. </b>Tissue section of human spleen using 1:200 dilution of Caspase-3 antibody (clone 31A1067). The staining was developed with HRP labeled anti-mouse IgG secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin. This Caspase 3 antibody generated primarily a specific cytoplasmic staining in a subset of spleenocytes with some nuclear signal in a few cells.

Simple Western lane view shows a specific band for Caspase-3 Antibody (31A1067) - (Pro and Active) in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230kDa separation system.

Caspase-3 was detected in immersion fixed paraffin-embedded sections of human bladder tissue using 1:50 dilution of mouse aCaspase-3 Antibody (31A1067) - (Pro and Active) (NB100-56708), for 1 hour at room temperature followed by anti-mouse IgG VisUCyte HRP polymer(VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue).

Paclitaxel in combination with MWE retarded tumor growth in a human bladder carcinoma TSGH 8301 xenograft model. The levels of total (t-PTEN) and phospho-PTEN (p-PTEN) and Caspase 3 in the tumor specimens were determined by Western blotting and then quantified using beta-actin as the protein loading control; the results are expressed as a percentage of the control. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep20417), licensed under a CC-BY license.

An intracellular stain was performed on NIH3T3 cells with Caspase-3 Antibody (31A1067) - (Pro and Active) Antibody NB100-56708AF700 (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 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 700.

<b>Biological Strategies Validation. </b>Lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with VP-16. PVDF membrane was probed with 0.1 ug/mL of mouse monoclonal Caspase-3 Antibody (31A1067) - (Pro and Active) (NB100-56708, Novus Biologicals) followed by 1:2000 dilution donkey anti-mouse IgG.

An intracellular stain was performed on HeLa cells with Caspase-3 Antibody (31A1067) - (Pro and Active) NB100-56708 (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, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems).

Western Blot: Mouse Monoclonal Caspase-3 Antibody (31A1067) - (Pro and Active) [IMGENEX: IMG-144A] [NB100-56708] - Whole cell lysates from MDA-MB-231, MCF-7, SUM149 and SUM159 cells were loaded with 50 ug/lane. 10% SDS-PAGE. Caspase-3 Antibody (NB100-56708) was used for primary antibody: 1:2000, 4℃, overnight. Image from a verified customer review.

Immunocytochemistry/ Immunofluorescence: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - rOPN administration influenced the interaction & balance between Beclin 1 & Caspase‐3 at 24 h after SAH. Double immunofluorescence staining of Caspase‐3 & Beclin 1 in Sham group, SAH + Vehicle group & SAH + rOPN group at 24 h after SAH induction. Sample size is 9, n = 3 per group. Localization of Caspase‐3 can be cytoplasmic & nuclear. Staining in the nucleus is considered to be an indication of active Caspase‐3. The dashed lines & the red box on brain slice images indicate the locations observed. Vehicle, phosphate‐buffered saline; Scale bar = 50 μm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31436915), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - PEM induces YIPF2 upregulation & apoptosis in NSCLC cells.a, b H1792 (a) & H1299 (b) NSCLC cells were treated with PEM at various concentrations (0–10.0 μM) for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2 & ACTB. c Overexpression of YIPF2 in H1299 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, CASP8, CASP3, PARP1 & ACTB. d Knockdown of YIPF2 expression by YIPF2–1 siRNA in A549 NSCLC cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, CASP8, CASP3, PARP1 & ACTB. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32303681), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - Lipofection of top broccoletti-miR candidates does not influence basal & induced apoptosis.(A) BxPc-3 & Bx-Gem cells were transfected as described in Figure 3. Seventy-two hours later, the cells were stained with Annexin V-PE & 7-AAD, followed by FACS analysis. The percentage of Annexin V- & 7-AAD-positive cells is shown. (B) Lipofected BxPc-3 & Bx-Gem cells were stained with an antibody specific for the proliferation marker Ki-67 (green) or the apoptosis marker cleaved fragment of caspase-3 (red), which indicates apoptosis. Representative images at ×100 magnification are shown. The percentage of Ki-67- or caspase-3-positive cells was counted in 18 visual fields, & the means ± SD are shown in the diagram below. (C) BxPc-3 & Bx-Gem were lipofected as described above, & at 24 h later, the cells were treated with gemcitabine (10 nM) or were left untreated. Ninety-six hours after gemcitabine treatment, viability was determined by MTT assay. The data are presented as the means ± SD (**P < 0.01). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32292571), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - PEM induces apoptosis of NSCLC cells via YIPF2-TNFRSF10B axis.a, b A549 (a) & H1792 (b) cells were treated with PEM at 5.0 μM for the indicated times (0, 6, 12, 24, 36 & 48 h). Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B & ACTB. c A549 cells were treated with doxorubicin (DOX) at various concentrations (0–2.0 μM) for 18 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B & ACTB. d Overexpression of YIPF2 in H1792 & H1299 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B & ACTB. e Knockdown of YIPF2 expression by YIPF2–1 siRNA in A549 & H1792 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B & ACTB. f Overexpression of YIPF2 in H1299 cells (left) or Knockdown of YIPF2 expression by YIPF2–1 siRNA in H1792 cells (right) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10A & ACTB. g Three A549 cell lines (Ctrl, YIPF2, YIPF2 + siTNFRSF10B) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B, CASP8, CASP3 & ACTB. h Three H1299 cell lines (Ctrl, siYIPF2–1, siYIPF2–1 + TNFRSF10B (short isoform)) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B, CASP8, CASP3 & ACTB. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32303681), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - Cell numbers positively stained for NeuN, DCX, PCNA & caspase-3 in the subgranular region of the dentate gyrus in WT & RanBP9−/− (KO) mice.(A), DAPI-stained brain sections to show the highlighted subgranular zone within the dentate gyrus region of the hippocampus used for cell counts shown in B. (B), Representative brain sections stained with anti-NeuN, anti-DCX, anti-PCNA, anti-capsase-3 & counter stained with DAPI. Cell counting revealed significantly decreased numbers of NeuN positive cells in RanBP9−/− (KO) brains (22%) compared to WT controls. However DCX, PCNA & caspase positive cell numbers were not significantly altered. In each group, n = 3, data presented as mean± SEM. **, p<0.01 by Student’s t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23840553), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - NFATc1 regulates DcR3 expression at a transcriptional level. (A,B) Immunoblot analysis of whole-cell lysates & quantitative real-time-PCR assaying relative DcR3 mRNA expression of ACHN & 769-P cells 24 h after treatment with cyclosporin A (CsA, 25 μM) or Tacrolimus (FK-506, 50 μM) (A); 48 h post transfection with NFATc1 or an empty vector control (neo) (B). Expression data were normalized to internal 18S rRNA expression (mean ± SEM; n=3; *p<0.05, **p<0.01, ***p<0.001; T-test). (C) Immunoblot analysis of cytoplasmic & nuclear fractions of ACHN & 769-P cells after treatment with LY294002 (50 μM), Everolimus (1 μM), or Cyclosporine A (25 μM). (D) Relative NFATc1–luciferase reporter activity of ACHN & 769-P cells 24 h post transfection with myrAkt or an empty vector control (neo) (mean ± SEM; n=3; *p<0.05, **p<0.01; T-test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24107265), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - rOPN administration elevated the expression of autophagy‐related proteins while suppressing apoptosis in rat brain at 24 h after SAH. The effects of rOPN on expression levels of (A) Beclin 1, mean ± SD is 1.016 ± 0.2262 in Sham group, 2.874 ± 1.147 in SAH + Vehicle group, 4.963 ± 2.05 in SAH + rOPN group, F = 15.52, (B) ATG5, mean ± SD is 0.8908 ± 0.2545 in Sham group, 2.332 ± 0.6431 in SAH + Vehicle group, 4.364 ± 1.309 in SAH + rOPN group, F = 25.02, (C) LC3, mean ± SD is 1 ± 0.1845 in Sham group, 1.755 ± 0.3017 in SAH + Vehicle group, 2.7 ± 0.7957 in SAH + rOPN group, F = 17.23, (D) Cleaved Caspase‐3, mean ± SD is 1.008 ± 0.186 in Sham group, 4.112 ± 1.528 in SAH + Vehicle group, 2.291 ± 0.6268 in SAH + rOPN group, F = 15.86, (E) Bax, mean ± SD is 1.006 ± 0.321 in Sham group, 37.47 ± 10.86 in SAH + Vehicle group, 23.83 ± 8.143 in SAH + rOPN group, F = 33.13, (F) Bcl‐2, mean ± SD is 1.005 ± 0.2736 in Sham group, 0.2309 ± 0.1257 in SAH + Vehicle group, 0.7843 ± 0.2278 in SAH + rOPN group, F = 20.1, in the left hemisphere of rat brain at 24 h after SAH. Sample size is 18, n = 6 per group. Data were presented as mean ± SD. *P < .05, ***P < .001 vs Sham group; #P < .05, ##P < .01 vs SAH + Vehicle group Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31436915), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free [NB100-56708] - RAB8 suppresses PEM-induced apoptosis of NSCLC cells by promoting the removing of TNFRSF10B from plasma membrane to cytoplasm.a, b Knockdown of RAB8 expression by RAB8 siRNA in H1792 (a) & A549 (b) cells in the presence or absence of PEM at 5.0 μM for 36 h. The surface expression of TNFRSF10B was confirmed by flow cytometry analyses. c Overexpression of RAB8 in H1299 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against RAB8, TNFRSF10B, CASP8, CASP3, PARP1 & ACTB. d Knockdown of RAB8 expression by RAB8 siRNA in H1792 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against RAB8, TNFRSF10B, CASP8, CASP3, PARP1 & ACTB. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32303681), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ch, ChHa, Ma

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ch, ChHa, Ma

Applications:

, , , , , , , , , , ,

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

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-144A

Alt Catalog# IMG-144A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-56708AF350
Alexa Fluor 405	NB100-56708AF405
Alexa Fluor 488	NB100-56708AF488
Alexa Fluor 532	NB100-56708AF532
Alexa Fluor 594	NB100-56708AF594
Alexa Fluor 647	NB100-56708AF647
Alexa Fluor 700	NB100-56708AF700
Alexa Fluor 750	NB100-56708AF750
Allophycocyanin	NB100-56708APC
Biotin	NB100-56708B
CoraFluor 1	NB100-56708CL1
DyLight 350	NB100-56708UV
DyLight 405	NB100-56708V
DyLight 488	NB100-56708G
DyLight 550	NB100-56708R
DyLight 594	NB100-56708DL594
DyLight 650	NB100-56708C
DyLight 680	NB100-56708FR
DyLight 755	NB100-56708IR
FITC	NB100-56708F
HRP	NB100-56708H
Janelia Fluor 525	NB100-56708JF525
Janelia Fluor 549	NB100-56708JF549
Janelia Fluor 646	NB100-56708JF646
Janelia Fluor 669	NB100-56708JF669
PE	NB100-56708PE
PerCP	NB100-56708PCP
mFluor Violet 450 SE	NB100-56708MFV450
mFluor Violet 500 SE	NB100-56708MFV500
mFluor Violet 610 SE	NB100-56708MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-56708SS
Azide and BSA Free	NBP2-33244

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NBP1-21502

Ferroportin/SLC40A1 Antibody - BSA Free

(7 Reviews)

(194 Publications)

0.1 ml

NBP1-21502

$Mouse Ferroportin/SLC40A1, 5-aminolevulinic acid synthase (ALAS), and ferritin light chain (FLC) proteins are increased in sickle mice overexpressing human wt-FHC. Proteins of subcellular fractions isolated from livers of wt-, ms-, and LRS-treated mice (n = 4) were run on a western blot (30 ug protein/lane) and immunostained for microsomal ferroportin. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fphar.2014.00079/abstract), licensed under a CC-BY license.$

HepG2 cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with Ferroportin/SLC40A1 Antibody at 5.0 ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (green) at a 1:500 dilution. Alpha tubulin (DM1A) [NB100-690] was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (red) at a 1:500 dilution. Nuclei were counterstained with DAPI (blue). Cells were imaged using a 40X objective.

Detection of Ferroportin/SLC40A1 protein in murine small intestinal section using Ferroportin/SLC40A1 Antibody at a dilution of 1:200. The antibody primarily developed a membranous staining pattern in the intestinal epithelial cells.

An intracellular stain was performed on HepG2 cells with Ferroportin/SLC40A1 Antibody NBP1-21502 (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 of human intestine lysate (Molecular weight: 62.5 KDa) using Ferroportin/SLC40A1 Antibody.

Western blot of bovine adult eye using Ferroportin/SLC40A1 Antibody. Image from verified customer review.

Duodenal mucosa samples from weanling pigs with iron replete or iron deficient status were probed using (NBP1-21502; 1:1000 dilution) and GAPDH (loading control). Image from verified customer review.

HepG2 clones overexpressing heparanase showed a reduction of hepcidin expression and indices of iron loading. Two stable clones of HepG2 cells transfected with pcDNA3.1-HPA (HPA3 and HPA6) were analyzed for hepcidin expression, BMP/SMAD signaling and indices of iron status. Western blot of Ferroportin/SLC40A1 and its densitometry. The values are expressed as-dCt (for HPA mRNA) or as fold change over the control (MOCK) (for hepcidin and Id1 mRNA). The images are representative from three different analyses. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371:journal.pone.0164183), licensed under a CC-BY license.

Immunostaining for Ferroportin/SLC40A1 shows uniform reactivity on the ciliary epithelium (panel 1). Fpn is expressed mainly in NPE cells (panel 2). H&E stained section shows orientation of the ciliary epithelium with respect to the iris and AH (panel 3). Fpn is expressed on corneal epithelium (panel 4) and endothelium (panel 5). Scale bar: 25 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29859760/) licensed under a CC-BY license.

Detection of Ferroportin/SLC40A1 protein in murine liver section using Ferroportin/SLC40A1 Antibody at a dilution of 1:200. The representative image shows intense staining in the cellular membranes, whereas, a relatively milder postivity was observed in the cytoplasm of hepatocytes.

Analysis using the PE conjugate of Ferroportin/SLC40A1 Antibody. Staining of Ferroportin in human B lymphocytes in peripheral blood using PE conjugated anti-Ferroportin/SLC40A1 antibody. Image from verified customer review.

An intracellular stain was performed on HepG2 cells with NBP1-21502AF647 (blue) and a matched isotype control (orange, NBP2-24893AF647). 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 647.

An intracellular stain was performed on Hek293 cells with NBP1-21502 (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).

$Mouse ferroportin, 5-aminolevulinic acid synthase (ALAS), and ferritin light chain (FLC) proteins are increased in sickle mice overexpressing human wt-FHC. Proteins of subcellular fractions isolated from livers of wt-, ms-, and LRS-treated mice (n = 4) were run on a western blot (30 ug protein/lane) and immunostained for (A) microsomal ferroportin.$

Immunohistochemistry-Paraffin: Rabbit Polyclonal Ferroportin/SLC40A1 Antibody [NBP1-21502] - Staining of mouse small intestines using Ferroportin/SLC40A1 Antibody. Tissue was fixed in formalin overnight and embedded in paraffin. ~5 micro sections on slides were dehydrated. Antigen retrieval was done using citrate buffer in rice steamer for 20mins. 1 hour blocking without permeabilization. Overnight primary ab incubation was done 1:200 (NBP1-21502) at 4C. 2-hr secondary antibody incubation was at room temp; mounting media with DAPI. Image from a verified customer review.

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Analysis of Cu & Fe content & metal-binding protein expression in wild-type & PrPC-null mouse total brain at different ages. (A) The graph shows the ratio of Cu & Fe levels in Prnp0/0 & Prnp+/+ brain samples (P1, P365 N = 4; P7, P30, P180 N = 6; P90 N = 5). (B) Representative Western blot images showing metal-binding protein levels in Prnp0/0 & Prnp+/+ brain samples (N = 4). The constant level of the housekeeping proteins ( beta -III Tubulin & beta -Actin) are also reported. (C) The graph shows the up- or down-regulation of protein expression in Prnp0/0 samples compared to Prnp+/+, i.e., (Prnp0/0 protein OD/housekeeping OD)/(Prnp+/+ protein OD/housekeeping OD). All error bars indicate SD; N = 4 minimum; *p 0.05 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28864822), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - SHP abrogates the BMP6 effect on iron metabolism through inhibition of hepcidin gene expression in mice.(a–d) C57/BL6 mice were injected with Ad-GFP (n = 4 per group, 5.9 × 109 pfu) or Ad-Flag-SHP (n = 5 per group, 5.9 × 109 pfu) via the tail-vein, & treated with Vehicle or BMP6 (500 μg/kg, i.p.) for 6 h at day 5 after the infection. (a) Serum iron level. (b) Q-PCR analysis showing hepcidin & SHP mRNA levels in liver. (c) Serum hepcidin levels. (d) Western blot analysis showing SMAD1/5/8 phosphorylation & SHP expression in liver. (e) Western blot analysis showing FPN expression in spleen (top). Graphical representation showing FPN expression (bottom). Data are presented as means ± SD. Arrows show locations of molecular weight markers. The western blot images were cropped with a grey cropping line. All gels for western blot analysis were run under the same experimental conditions. **P < 0.01, ***P < 0.001 by two-tailed Student t-test. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep34630), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Upregulation of H3K4me3 increases iron exporter Fpn1 & exhibits neuroprotection.a, b CPI-455 treatment (24 h) increased the protein level of H3K4me3 & Fpn1 in SH-SY5Y cells. c MTT assay showed CPI-455 showed neuroprotection against the toxicity of 6-OHDA in SH-SY5Y cells. d–g SH-SY5Y cells were infected by AAV-KDM5C-silencing virus, & KDM5C was significantly decreased at the mRNA level as reflected by RT-PCR d & at the protein level by Western blot e, f. In addition, Fpn1 & H3K4me3 were increased in AAV-KDM5C-silencing virus infected cells e, f. Moreover, g silencing KDM5C showed a slight neuroprotection against 6-OHDA-induced PD model in SH-SY5Y cells. *P < 0.05; **P < 0.01; ***P < 0.001. Data are presented as mean ± SEM. n = 3–4 for each group in Western blot & n = 5 for each group in MTT assay. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33116116), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - SHP deficiency alters hepcidin gene expression in liver of HID mice.(a–i) WT & SHP KO mice (n = 5 per group) were fed with high-iron diet (HID, 8 g/kg) for 3 weeks. (a) Serum iron level. (b) Hepcidin mRNA level in liver. (c) Hepcidin expression in mouse liver. IHC was performed using an antibody against hepcidin. Scale bar shows 50 μm. (d) Serum hepcidin level. (e) Western blot analysis (left panel) showing hepatic SHP & splenic FPN expression & graphical representation (right panel) showing splenic FPN expression. (f) Splenic iron level. (g) Perls’ prussian blue staining in spleen. Scale bar shows 200 μm. (h) BMP6 & BMP9 mRNA levels in liver. (i) Western blot analysis (top) & graphical representation (bottom) showing SMAD1/5/8 phosphorylation in liver. The grouping of the images is from different parts of the same gel. Data are presented as means ± SD. Arrows show locations of molecular weight markers. The experiment was repeated on a minimum of three separate occasions. The western blot images were cropped with a grey cropping line. All gels for western blot analysis were run under the same experimental conditions. *P < 0.05, **P < 0.01, ***P < 0.001 by two-tailed Student t-test. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep34630), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Differential expression of proteins of iron metabolism in CCA cells. The CCA4, CCLP1 & HUCCT1 human CCA-derived cell lines were cultured as adherent monolayers (MON) or in 3D sphere conditions (SPH). Panel a. Top left, representative immunoblot analysis. Cell extracts were reacted with antibodies against transferrin receptor (TfR1), ferroportin (FPN), ferritin H subunit (FtH) & vinculin. Cropped blots are displayed. The original full blot images can be found in Supplementary Information. The graphs show densitometric quantification of immunoblot analyses. The values were normalized to vinculin & expressed as a fraction of respective MON cells normalized to 1. Mean values ± SEM (n = 6), *p ≤ 0.05, **p ≤ 0.01 vs control MON for each cell line. Panel b. RNA bandshift analysis of IRP activity. Cytoplasmic extracts were incubated with a 32P-labeled iron-responsive element (IRE) probe & RNA-protein complexes separated on non-denaturing polyacrylamide gels. On the left a representative autoradiogram is shown. A cropped gel is displayed. The original full gel image can be found in Supplementary Information. The graph on the right shows the densitometric quantification of IRPs bands by direct nuclear counting, as described in Materials & Methods; mean percentages ± SEM of control values (n = 6),, **p ≤ 0.01, ***p< 0.001 vs control MON for each cell line. Panel c. TfR1 & FPN mRNA levels were measured by quantitative RT-PCR. Samples were analyzed in triplicate, normalized to the housekeeping gene 18 S & expressed as percentage of respective MON cells normalized to 1. Mean values ± SEM (n = 6), **p ≤ 0.01 vs control MON for each cell line. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29247214), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Iron (55Fe) transport across the apical & basal borders. (a) Apical transport (uptake) of 55Fe in response to intracellular/basal ascorbate (150 μM) was increased compared to no ascorbate. The cells were incubated with ascorbate in the basal chamber for 24 h. Values are means of 10 samples ±SD. The difference between treatments was significant (p = 0.04). (b) Basolateral transport of iron (as 55Fe) in response to intracellular/basal ascorbate (150 μM) was increased compared to no ascorbate. The cells were incubated with ascorbate in the basal chamber for 24 h. Values are means of 10 samples ±SD. The difference between treatments was significant (p = 0.03). (c) Western Blot of cells treated with ascorbate (150 μM). Lane 1: Control cells (no treatment). Lane 2: Basal ascorbate at 150 μM; these were the ferroportin levels before iron addition in (a). Lane 3: Ferroportin levels 22 h after the iron addition (20 μM for 2 h) in (a). Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/2072-6643/6/1/249), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Metformin rescues BMP6-mediated alteration of iron metabolism in mice.(a–e) C57/BL6 mice (n = 4 per group) were treated with BMP6 (500 μg/kg, i.p.) & metformin (200 mg/kg, p.o.). (a) Serum iron level. (b) Q-PCR analysis showing hepcidin & SHP mRNA levels in liver. (c) Serum hepcidin levels. (d) Western blot analysis showing SMAD1/5/8 phosphorylation & FPN in liver. (e) Western blot analysis (top) & graphical representation (bottom) showing FPN expression in spleen. (f) Schematic diagram of SHP-mediated inhibition of BMP6-SMADs pathway. Data are presented as means ± SD. Arrows show locations of molecular weight markers. The western blot images were cropped with a grey cropping line. All gels for western blot analysis were run under the same experimental conditions. *P < 0.05, **P < 0.01, ***P < 0.001 by two-tailed Student t-test. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep34630), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Comparison of Cu, Fe, & metal-binding protein expression levels in wild-type & PrPC-null mouse spleen at different ages. (A) The graph shows the ratio of Cu & Fe levels in Prnp0/0 & Prnp+/+ spleen samples (P15 N = 3; P30 N = 4; P90, P180 N = 6; P365 N = 5). (B) The graph shows the weight of spleen extracted from Prnp0/0 & Prnp+/+ mice; N = 4. (C) Representative Western blot images showing metal-binding protein levels in Prnp0/0 & Prnp+/+ spleen samples. The constant level of the housekeeping protein ( beta -Actin) are also reported. (D) The graph shows the up- or down-regulation of protein expression in Prnp0/0 samples compared to Prnp+/+, i.e., (Prnp0/0 protein OD/housekeeping OD)/ (Prnp+/+ protein OD/housekeeping OD); N = 4. All error bars indicate SD; *p < 0.05; **p < 0.01; ***p < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27729845), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Htt elimination disrupts brain iron homeostasis.(A) Representative photographs of immunohistochemical staining for ferritin light chain (Ft). Left panels: cerebella of 21mo CTL noTM & 18mo cKO TM@3mo; middle panels: thalami of 12mo CTL TM@9mo & 12mo cKO TM@9mo; right panels: striata of 21mo CTL noTM & 18mo cKO TM@3mo. Note that Ft levels are already extremely reduced in the thalamus 3 months after Htt elimination (middle panels), & that Ft expression is practically abolished in the cerebellum in the absence of Htt (left panels). (B) Representative western blots of Tfr & Fpn protein expression in 10 month-old brains from CTL & cKO mice TM-treated at 6 months of age. Antibody against actin was used as internal control for loading. Note that Tfr & Fpn levels are increased in TM-treated cKO compared to controls. (C) Quantification of Tfr & Fpn expression levels. Western blots of total protein extracts from 10 month-old brains CTL (n = 5) & cKO (n = 5) TM-treated at 6 months of age were probed with anti-Tfr or anti-Fpn, stripped & re-probed with anti-actin antibody. Bands intensities were quantitated using Image J. Tfr & Fpn levels were normalized over actin levels. Values represent mean relative to controls ± SD (***P<0.001, Student’s t-test). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pgen.1006846), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - GW501516 activation of PPARδ regulates 6-OHDA-triggered expression of level of DMT1 & FPN1 protein, & DMT1 mRNA, but not FPN1 mRNA. (A) Cells were treated with 6-OHDA for the indicated durations. (B,C) Cells pretreated with DMSO or GW501516 for 8 h were incubated with or without 6-ODHA for 16 h. (D) Cells were treated with GW501516 for the indicated durations. (E,F) Cells pretreated with DMSO or GW501516 for 8 h were incubated with or without 6-ODHA for 16 h. Total RNA & protein were extracted, & mRNA & protein levels were analyzed by real-time PCR (A,B,D,E) & Western blot (C,F), respectively. RPS18 & alpha -tubulin were used as internal controls for real-time PCR & Western blot, respectively. Results are expressed as means ± SE (n = 3). An image analyzer was used to quantify band intensity of Western blot, & the ratio of protein to alpha -tubulin is indicated above each lane. * p < 0.05, ** p < 0.01 relative to the untreated group; #p < 0.05, ##p < 0.01 relative to the 6-OHDA-treated group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35624674), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Fisetin protects against DOX-induced ferroptosis via regulating Nrf2 in rats. (A). Western blot results of Nrf2 & Keap1, HO-1, FTH1, FTL, FPN, & TfR1 protein in control & different drug-treated rat heart tissues. (B). Quantification of (A). (C). Expression of Nrf2 was detected by immunohistochemistry (IHC) (Representative images, 200X & 400X, Scale bar = 100 & 50 μm, n = 6 rats per groups) in cardiac tissue of each group. The values are presented as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001; NS, no significance; DOX, doxorubicin. Nrf2, nuclear factor erythroid 2-related factor 2; Keap1, Kelch-like ECH-associated protein 1; HO-1, heme oxygenase-1; FTH1, ferritin heavy chain 1; FTL, ferritin light chain; FPN, ferroportin; TfR1, transferrin receptor 1. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35273493), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Distribution of TfR & Fpn in the anterior segment of bovine eye:(A) The reaction for TfR is evident on the ciliary epithelium facing the AH (panel 1). TfR is localized on the apical membrane & intracellular vesicles of NPE cells as in the human sample (panel 2). The corneal epithelium does not react for TfR (panel 3), while the corneal endothelium shows a distinct reaction on the plasma membrane (panel 4). H&E staining shows multiple layers of corneal epithelium, the stroma, & a single layer of corneal endothelium (panel 5). Scale bar: 25 μm. (B) Fpn is expressed on the ciliary epithelium & the iris (panel 1). The expression of Fpn is limited to NPE cells as in the human sample (panel 2). H&E staining shows the orientation of ciliary epithelium with respect to the iris & AH (panel 3). Unlike human samples, the bovine corneal epithelium did not react for Fpn (panel 4). However, corneal endothelium showed strong reactivity for Fpn (panel 5). Scale bar: 25 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29859760), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - GW501516 PPARδ activation suppresses 6-OHDA-induced expression of IRP1 in SH-SY5Y cells. (A,B) Cells were treated with 6-OHDA (A) or GW501516 (B) for the indicated durations. (C) Cells stably expressing shRNA targeting scrambled sequences or PPARδ were pretreated with DMSO or GW501516 for 8 h & subsequently incubated in the presence or absence of 6-ODHA for 16 h. (D,E) Cells pretreated with DMSO or GW501516 for 8 h were incubated with or without 6-ODHA for 16 h. Total RNA & protein were extracted, & levels of mRNA & protein were analyzed by real-time PCR (A–D) & Western blot (E). The immunoblots are separate from those shown in Figure 4C,F. RPS18 & alpha -tubulin were used as internal controls for real-time PCR & Western blot, respectively. Results are expressed as means of triplicate ± SE (A–D). * p < 0.05, ** p < 0.01 relative to untreated group; #p < 0.05 relative to 6-OHDA-treated group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35624674), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Ferroportin/SLC40A1 Antibody - BSA Free [NBP1-21502] - Expression of iron modulating proteins in the cornea & ciliary body.(A) Probing of lysates from bovine cornea (Cor) & CB for Cp, TfR, Tf, & ferritin shows the expression of all of these proteins in both samples (lanes 1–4). Deglycosylation results in faster migration of Cp, TfR, & Tf on SDS-PAGE, indicating the presence of glycans (lanes 2 & 4). AH & the vitreous show abundant presence of Cp & Tf, both of which migrate faster upon deglycosylation (lanes 7 & 9). No reactivity for TfR or ferritin is detected in these samples (lanes 6–9). Human brain lysate was processed in parallel as a positive control (lane 5). Gapdh served as a loading control. (Cor: cornea; Ft: ferritin). (B) Relative distribution of iron modulating proteins within each tissue shows higher expression of TfR relative to Cp & ferritin in the cornea, & higher levels of Cp relative to the TfR & ferritin in the CB. (C) Quantitative comparison of protein expression by densitometry shows significantly higher levels of ferritin & Cp, & lower levels of TfR in the CB relative to the cornea. All values were normalized to Gapdh that provided the loading control. Values represent fold change ± SEM of the indicated n. (D) Probing of Western blots of bovine cornea & CB for Fpn revealed increased expression of Fpn (3.2 fold) in CB relative to the cornea (lanes 1–4). Lysates from human brain & bovine retina were analyzed in parallel as controls (lanes 5 & 6). Gapdh served as a loading control. (E) Quantification by densitometry shows 3.2 fold higher levels of Fpn in the CB relative to the cornea. Values are mean + SEM of the indicated n. **p < 0.01. The full images of the cropped blots have been provided in the Supplementary Data. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29859760), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-21502AF350
Alexa Fluor 405	NBP1-21502AF405
Alexa Fluor 488	NBP1-21502AF488
Alexa Fluor 532	NBP1-21502AF532
Alexa Fluor 594	NBP1-21502AF594
Alexa Fluor 647	NBP1-21502AF647
Alexa Fluor 700	NBP1-21502AF700
Alexa Fluor 750	NBP1-21502AF750
Allophycocyanin	NBP1-21502APC
Allophycocyanin/Cy7	NBP1-21502APCCY7
Biotin	NBP1-21502B
CoraFluor 1	NBP1-21502CL1
DyLight 350	NBP1-21502UV
DyLight 405	NBP1-21502V
DyLight 488	NBP1-21502G
DyLight 550	NBP1-21502R
DyLight 594	NBP1-21502DL594
DyLight 650	NBP1-21502C
DyLight 680	NBP1-21502FR
DyLight 755	NBP1-21502IR
FITC	NBP1-21502F
HRP	NBP1-21502H
Janelia Fluor 525	NBP1-21502JF525
Janelia Fluor 549	NBP1-21502JF549
Janelia Fluor 646	NBP1-21502JF646
Janelia Fluor 669	NBP1-21502JF669
PE	NBP1-21502PE
PE/Atto594	NBP1-21502PEATT594
PE/Cy5.5	NBP1-21502PECY55
PE/Cy7	NBP1-21502PECY7
PerCP	NBP1-21502PCP
mFluor Violet 450 SE	NBP1-21502MFV450
mFluor Violet 500 SE	NBP1-21502MFV500
mFluor Violet 610 SE	NBP1-21502MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP1-21502SS

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NB400-132

ABCG1 Antibody - BSA Free

(2 Reviews)

(186 Publications)

0.1 ml

NB400-132

Flow (Intracellular): ABCG1 Antibody - BSA Free [NB400-132] - RAW 246.7 cells were either untreated (A) or serum starved for 24 hours, and then treated with 1uM TO9 for 24 hours (B). An intracellular stain was performed with NB400-132 (blue) and a matched isotype control NB810-56910 (orange). Cells were fixed with 4% paraformaldehyde, following fixation, cells were permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature, followed by Goat Anti-Rabbit Dylight 550-conjugated antibody.

<b>Biological Strategies Validation. </b>Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - Cells were pretreated with 5 uM F12511 or 5 uM MG132 for 30 min and then exposed to 5 or 50 uM 24S-OHC for 6 h. Whole-cell lysates were immunoblotted with antibodies specific for ABCG1 or beta-actin. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31285856/) licensed under a CC-BY license.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - Detection of ABCG1 in J774 mouse macrophages. Image provided by Dr. Angeliki Chroni of the National Centre for Scientific Research Demokritos.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - Raw 264.7 (A), and Raw 264.7 + 10uM T09 18 hr (B) cell lysate using ABCG1 antibody at 2 ug/mL.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - Detection of ABCG1 in mouse peritoneal macrophages. Lane 1: untreated mouse peritoneal macrophages, Lane 2: 22-(R)-hydrocholesterol treated macrophages, Lane 3: T0901713 treated macrophages, Lane 4: T091713 treated macrophages.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - SAA has little impact on the impairment in biliary and fecal excretion of macrophage-derived 3H-cholesterol during endotoxemia. WT and SAAKO mice were injected subcutaneously with PBS or 0.8 ug/g LPS as indicated, followed 4 hr later by intraperitoneal injection of 3H-cholesterol-labeled J774 macrophages. ABCG1 in liver lysates (40 ug protein) was identified by immunoblotting and quantified by densitometry. Results from each of the 3 experiments (n = 4-5 per group per experiment) were expressed relative to PBS-injected WT mice after normalization to beta-actin. A representative western blot is shown. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/jl/2013/283486/), licensed under a CC-BY license.

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

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

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

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

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

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - 6‐Dihydroparadol increases A) ABCA1 & B) ABCG1 protein stability. THP‐1 cells were differentiated as described in Figure 2. Then cells were loaded with unlabeled cholesterol & treated with 6‐dihydroparadol (10 μm), or solvent vehicle (0.1% DMSO, control) for 24 h. Cells were lysed at different time points (0, 1, 2, 3, 4, & 6 h) after treatment with the protein synthesis inhibitor cycloheximide (CHX, 100 μm). The protein levels of both ABCA1 & ABCG1 were detected by Western blot analysis. The data points represent mean ± SD from three independent experiments. *p < 0.05 & **p < 0.01 versus control at the same time point (determined by Student's t‐test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29802792), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ABCG1 Antibody - BSA Free [NB400-132] - Evodiamine enhances apo A1-mediated ChE from THP-1 macrophages & increases ABCA1 protein level. (a) Differentiated THP-1 cells were loaded with [3H]-cholesterol together with the indicated treatments for 24 h. On the next day, the cells were washed twice with PBS & incubated with the same compounds [solvent vehicle control (Veh; ≤0.1% DMSO), evodiamine (1–20 μM), & the PPAR gamma agonist pioglitazone (10 μM) as positive control] with or without 10 µg/mL apo A1. Extracellular as well as intracellular radioactivities were quantified with scintillation counter. Differentiated THP-1-derived macrophages were treated with solvent vehicle control (Veh; ≤0.1% DMSO), evodiamine (10 μM), & the PPAR gamma agonist pioglitazone (10 μM) as positive control. After 24 h incubation, the cells were lysed & 20 μg protein was resolved via SDS-PAGE. Immunodetection was performed with antibodies against the indicated proteins, ABCA1 (b), ABCG1 (c), & SR-B1 (d), & visualized by chemiluminescence detection. All experiments were performed at least three times & data are presented as means ± S.D. vs. solvent vehicle control, *p < 0.05, **p < 0.01, ***p < 0.001, n.s. no significance (ANOVA/Bonferroni). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30038271), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, ChHa, Ha, Pm, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, ChHa, Ha, Pm, Rb

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-132AF350
Alexa Fluor 405	NB400-132AF405
Alexa Fluor 488	NB400-132AF488
Alexa Fluor 532	NB400-132AF532
Alexa Fluor 594	NB400-132AF594
Alexa Fluor 647	NB400-132AF647
Alexa Fluor 700	NB400-132AF700
Alexa Fluor 750	NB400-132AF750
Allophycocyanin	NB400-132APC
Allophycocyanin/Cy7	NB400-132APCCY7
Biotin	NB400-132B
CoraFluor 1	NB400-132CL1
DyLight 350	NB400-132UV
DyLight 405	NB400-132V
DyLight 488	NB400-132G
DyLight 550	NB400-132R
DyLight 594	NB400-132DL594
DyLight 650	NB400-132C
DyLight 680	NB400-132FR
DyLight 755	NB400-132IR
FITC	NB400-132F
HRP	NB400-132H
Janelia Fluor 525	NB400-132JF525
Janelia Fluor 549	NB400-132JF549
Janelia Fluor 646	NB400-132JF646
Janelia Fluor 669	NB400-132JF669
PE	NB400-132PE
PE/Atto594	NB400-132PEATT594
PE/Cy5.5	NB400-132PECY55
PE/Cy7	NB400-132PECY7
PerCP	NB400-132PCP
mFluor Violet 450 SE	NB400-132MFV450
mFluor Violet 500 SE	NB400-132MFV500
mFluor Violet 610 SE	NB400-132MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-132SS

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NB100-123

HIF-1 alpha Antibody (H1alpha67)

(9 Reviews)

(165 Publications)

0.1 ml

NB100-123

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - MLN4924 induces accumulation of HIF1a in a time dependent manner. Cells were treated with 0.1uM MLN4924 for indicated time periods, followed by IB with indicated antibodies. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2012125) licensed under a CC-BY license.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Analysis of HIF-1 alpha in human lung tissue. Image courtesy of product review by Aneta Gandjeva.

Immunoprecipitation: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - MG132 attenuates the inhibitory effect of interleukin-1beta on HIF-1/AM axis. U87MG cells were incubated for 2 h in hypoxia (1% O2) with or without interleukin-1beta (10 ng/ml). The proteasome inhibitor MG132 was added 30 min in advance to prevent ubiquitinated HIF-1alpha from proteasomal degradation. HIF-1alpha was precipitated by anti-HIF-1alpha antibody. Ubiquitin was detected by immunoblotting. Non-specific mouse IgG was used as a negative control. The results are representative for three independent experiments Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2013562) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Analysis using the HRP conjugate of NB100-123. Detection of HIF-1 alpha in cobalt chloride treated/untreated COS-7 nuclear extracts using NB100-123.

$<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Analysis using the HRP conjugate of NB100-123. On day 1, MEF cells (+/+,-/-), were grown on 15cm dish (2x10 to the 6th cells). On day 2, cells were exposed to hypoxia for 4hrs. Cells were washed with ice cold PBS twice and whole cell protein was extracted extracted with RIPA buffer fortified with protease. Upon quantification, 100ug of protein was fractionated on 7% polyacralymide gel. Gel was transferred overnight onto nitrocellulose membrane. The membrane was probed with HIF-1 alpha monoclonal antibody at a 1:500 dilution (NB100-123). The secondary antibody was conjugated with HRP and was used at a 1:2500 dilution. Photo courtesy of Dr. Gregg Semenza, Johns Hopkins University.$

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Dapagliflozin protects against metabolic switch by inhibition of HIF-1 alpha. Representative images of PTCs incubated with 10 umol/l of molidustat for 6 h with immunofluorescent staining for HIF-1 alpha. Scale bar = 10 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33187129/) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Immunohistochemical detection of HIF-1alpha protein in human tissues. The nature of the tissue is indicated on top of each figure. Original magnifications are as follows: A, x400; B, x400; C, x100; D, x100; E, x100; F, x100; G, x40; H, x40. Image collected and cropped by CiteAb from the following publication (//bmcgenet.biomedcentral.com/articles/10.1186/1471-2156-5-27), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Staining in pig endothelial cells under hypoxia condition using NB100-123. Image from verified customer review.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Staining on pig tissue (brown) using NB100-123. Image from verified customer review.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Staining of biotin conjugated HIF-1 alpha (NB100-123B) in human kidney.

Immunoprecipitation: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Immunoprecipitation of HIF-1alpha using NB100-123 in COS7 CoCl2 treated lysate. Heavy and light chains are also detected. Image using the HRP form of this antibody.

Immunohistological and western blot analyses of HIF-1 alpha .(A) PAB liver stained by HIF-1 alpha (original magnification, ×20). (B) Sham-operated control liver stained by HIF-1 alpha (original magnification, ×20). (C) Enlarged view of the boxed area in the Image (A) (original magnification, ×100). (D) Western blot analysis of HIF-1 alpha (mean ± SEM), * P<0.05. HIF-1 alpha : hypoxia-inducible factor-1 alpha ; PAB: pulmonary artery banding.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: Mouse Monoclonal HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Images of HIF-1 alpha localization (red) in CTR and CoCl2-treated oAECs after 24 h of culture. Image from a verified customer review.

Western Blot: Mouse Monoclonal HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Image of HIF-1 alpha activation in AECs control cells (CTR) and after 24h of 10 and 50µM of CoCl2-treatment; HIF-1 alpha time course at shorter CoCl2 incubation times. Image from a verified customer review.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Immunohistochemical detection of HIF-1 alpha protein in human tissues. The nature of the tissue is indicated on top of each figure. Original magnifications are as follows: A, ×400; B, ×400; C, ×100; D, ×100; E, ×100; F, ×100; G, ×40; H, ×40. Image collected & cropped by CiteAb from the following publication (//bmcgenet.biomedcentral.com/articles/10.1186/1471-2156-5-27), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Immunohistological & western blot analyses of HIF-1 alpha .(A) PAB liver stained by HIF-1 alpha (original magnification, ×20). (B) Sham-operated control liver stained by HIF-1 alpha (original magnification, ×20). (C) Enlarged view of the boxed area in the Image (A) (original magnification, ×100). (D) Western blot analysis of HIF-1 alpha (mean ± SEM), * P<0.05. HIF-1 alpha : hypoxia-inducible factor-1 alpha ; PAB: pulmonary artery banding. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26863419), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Analysis of CBF treatment in HCT116 implanted mouse models & tumour tissue samples. (a) Tumour size versus CBF treatment. The lowest tumour growth rate was found in i.p. group during CBF treatment. (b) Analysis of HIF-1 alpha mRNA level in tumour tissues. There was a significant upregulated HIF-1 alpha level in i.p. group. Results are means with standard errors from 4 replicates. (c) Inhibition of HIF-1 alpha nuclear translocation in the i.t. sample. CBF strongly inhibited HIF-1 alpha nuclear accumulation in the tumour of xenografts implanted with HCT116 cells. Scale bars equal 10 μm. White arrows: HIF-1 alpha in the cytosol. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23818933), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - CycT alleviates tumor hypoxia. The effects of CycT on the levels of exogenous hypoxia-marker pimonidazole labeling (A), the levels of hypoxia-inducible CA9 enzyme (B), & the levels of hypoxia-inducible factor HIF1 alpha (C) in orthotopic tumor xenografts. Scale bar: Montage, 1 mm; 10X, 20 μm. Data are plotted as mean ± SEM. For statistical analysis, the levels in treated tumors were compared to the levels in control tumors with a Welch 2-sample t-test. **p-value < 0.005. IHC images are representative of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30723259), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Schematic demonstration of the three HIF1 alpha antibodies used in this study & their epitopes (Figs. 2a & b). Ab1 is a polyclonal antibody raised against N-domain of wild type HIF1 alpha & does not recognize HIF1 alpha 1.2 due to it's different N-terminal part (Fig. 2a). Ab2 & Ab3 are monoclonal & polyclonal antibodies, respectively, with epitopes in the common parts of HIF1 alpha & HIF1 alpha 1.2 (Fig.2a & b). Immunohistochemical analysis performed on thin adjacent sections of NE-differentiated prostate cancer using the HIF1 alpha antibodies Ab1 (Fig. 2c), Ab 2 (Fig. 2d), Ab 3 (Fig. 2e) & HIF1 beta antibody (Fig. 2f). Immunopositivity was detected for HIF1 alpha with Ab2 & Ab3 while HIF1 alpha Ab1 produced no detectable staining. HIF1 beta was also positive in adjacent section (Fig. 2f). Double staining of chromogranin A & androgen receptor antigens on adjacent sections (Fig. 2g) showed immunopositivity for chromogranin A (Fast red). Androgen receptor antibody (DAB, brown) produced no staining. Immunostaining of benign prostate tissue with HIF1 alpha Ab3 showed immunopositivity in NE-like cells of benign prostate tissue (Fig. 2h). In addition, HIF1 beta antibody recognized benign NE-like cells in benign prostate hyperplasia (Fig. 2i). Double staining with HIF1 alpha Ab3 & HIF1 beta (Fig. 2j) shows co-localization of the two proteins in NE-like cells of benign prostate hyperplasia (HIF1 alpha Ab3 red stain, HIF1 beta brown stain). Panels a, b, c, d, e, f & g: 40× objective. Panels h, i & j: 60× objective. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20663134), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - GAD65/67-positive neurons expressed HIF-1 alpha under hypoxic conditions. A) HIF-1 alpha expression co-localized with GAD65/67-ir in neurons exposed to hypoxia when compared to normoxia (upper panel) or GAD65/67-negative neurons in hypoxia (bottom panel, open arrow). B) Quantification shows the percentage of HIF-1 alpha -expressing GAD65/67-positive neurons after hypoxia in vitro (mean ± SD; from n = 6 cultures). C)In vivo immunostaining illustrates HIF-1 alpha -positive (bottom panel, solid arrow) & HIF-1 alpha -negative (bottom panel, open arrow) in GAD65/67-ir neurons in the ipsilateral region, whereas the contralateral region shows no HIF-1 alpha staining in GAD65/67-ir neurons (see Figure 1 for region selection). Scale bars, 10 μm (A); 20 μm (B). Image collected & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/2051-5960-2-51), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Levels of GSH in cortical neurons exposed to hypoxia. A) During hypoxia, GSH increased in MAP2-ir neurons with round somata (bottom panel, open arrow) & decreased in neurons with pyramidal-like morphology (upper panel, solid arrow). B) GSH increased in a subset of GAD65/67–ir neurons exposed to hypoxia (bottom panel, open arrow). C) Elevated HIF-1 alpha expression in GAD65/67–ir neurons containing high levels of GSH when exposed to hypoxia (middle panel). BSO treatment decreased HIF-1 alpha expression in GAD65/67-positive neurons during hypoxia. D) Percentages of HIF-1 alpha -expressing GAD65/67-ir neurons expressing a high level of GSH & GAD65/67-ir neurons expressing a low levels of GSH & HIF-1 alpha (mean ± SD; n = 3). The “low” level referred to the level of GSH in normoxic GAD65/67 neurons that was normalized to 1. The “high” level referred to the elevated GSH level in GAD65/67 neurons with a mean of 1.40 ± 0.10. E) Total MCB-GSH intensity in GAD65/67-positive neurons with & without BSO after hypoxia (mean ± SD; 8-10 neurons quantified from each experiment, n = 3 independent cultures). Scale bars, 20 μm. Image collected & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/2051-5960-2-51), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Results of in situ hybridization & immunohistochemistry on thin adjacent section to detect expression of HIF-1 alpha 1.2 mRNA & HIF1 alpha protein in malignant & benign prostate tissue. In situ hybridization (antisense probe, Fig. 3a) & immunostaining with HIF1 alpha Ab2 (Fig. 3c) on thin adjacent sections of NE-differentiated prostate adenocarcinoma showed co-localization of HIF1 alpha 1.2 transcript & HIF-1 alpha protein. Incubation with sense probe did not generate any detectable hybridization signals (Fig. 3b). Both In situ hybridization (Fig. 3d antisense) & HIF-1 alpha Ab2 immunostaining (Fig. 3f) were negative in non-NE-differentiated prostate adenocarcinoma. In situ hybridization with sense probe performed on non-NE-differentiated prostate cancer (Fig. 3e) was negative. In situ hybridizering on benign prostate tissue showed HIF1 alpha 1.2 transcript in NE-like cells of benign prostate tissue (Fig. 3g, ↗). The sense probe on thin adjacent section generated no signals (Fig. 3h,↗). Furthermore, co-localization of HIF1 alpha 1.2 transcript (Fig. 3i,↗,◢,↖) & HIF1 alpha protein, detected with HIF1 alpha Ab3 (Fig. 3j,↗,◢,↖) was also shown in NE-like cells of benign prostate tissue. Panels a, b, c, d, e, f, g, h, i, j: 40× objective. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20663134), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - HX-induced Sirt1 expression in white matter OPCs requires HIF1 alpha .(a) HIF1 alpha stabilization of Sirt1 transcript expression in OPCs as revealed by representative RT–PCR represents higher level of Sirt1 mRNA in VHL cKO mice. GDPDH mRNA serves as a control. Mean±s.e.m., n=3 brains for each group. (b,c) Representative western blot demonstrates a transient increase of HIF1 alpha expression in HX white matter at P18 with no significant effect at P11 (P=0.7955) & P45 (P=0.7333). Histograms show mean±s.e.m. (d,e) Graphs represent the percentages of Sirt1+Ki67+ & NG2+Ki67+ cells after HX white matter in WT & HIF1 alpha KO mice. Number in parentheses within bar indicates number of samples (n=4 brains per group & per genotype; ****P<0.0001, one-way analysis of variance, Bonferroni post hoc test, mean±s.e.m.). (f,g) Western blot demonstrates no increase (P=0.8231) in Sirt1 & HIF1 alpha expression in white matter of Hif1 alpha KO mice. Actin was used as loading control (mean±s.e.m.; n=3 brains for each experiment, group & genotype). Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms13866), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - HIF-1 alpha expression in primary cortical neurons exposed to hypoxia/ischemia. A & A’) Neurons were double-stained for HIF-1 alpha & MAP2 in the presence of 5 & 25 mM glucose with & without hypoxia. HIF-1 alpha expression in the somata was observed in cells with interneuron-like morphology after hypoxia. B) CoCl2 (0.3 mM) induced HIF-1 alpha expression in cells with interneuron-like morphology. C) Quantification represents the increase in HIF-1 alpha –ir staining (mean ± SD; 5-10 neurons quantified from each experiment, n = 3 experiments). D)In vivo brain slice shows a similar pattern of positive HIF-1 alpha -ir in round soma (open arrow) & negative in neurons with pyramidal-like morphology (solid arrow) in the ipsilateral side. *p < 0.05, compared with normoxia (25 mM glucose), #p < 0.05, compared with hypoxia (25 mM glucose). Scale bars, 20 μm (A, A’, B); 10 μm (D). Image collected & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/2051-5960-2-51), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Av, Bv, Ca, Ma, Pm, Rb, Sh

Species Glossary

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

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	NB100-123AF350
Alexa Fluor 405	NB100-123AF405
Alexa Fluor 488	NB100-123AF488
Alexa Fluor 532	NB100-123AF532
Alexa Fluor 594	NB100-123AF594
Alexa Fluor 647	NB100-123AF647
Alexa Fluor 700	NB100-123AF700
Alexa Fluor 750	NB100-123AF750
Allophycocyanin	NB100-123APC
Biotin	NB100-123B
CoraFluor 1	NB100-123CL1
DyLight 350	NB100-123UV
DyLight 405	NB100-123V
DyLight 488	NB100-123G
DyLight 550	NB100-123R
DyLight 594	NB100-123DL594
DyLight 650	NB100-123C
DyLight 680	NB100-123FR
DyLight 755	NB100-123IR
FITC	NB100-123F
HRP	NB100-123H
Janelia Fluor 525	NB100-123JF525
Janelia Fluor 549	NB100-123JF549
Janelia Fluor 646	NB100-123JF646
Janelia Fluor 669	NB100-123JF669
PE	NB100-123PE
PerCP	NB100-123PCP
mFluor Violet 450 SE	NB100-123MFV450
mFluor Violet 500 SE	NB100-123MFV500
mFluor Violet 610 SE	NB100-123MFV610

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NB100-56566

TLR4 Antibody (76B357.1) - BSA Free

(5 Reviews)

(148 Publications)

0.1 mg

NB100-56566

Immunohistochemistry: TLR4 Antibody (76B357.1) [NB100-56566] - Pericryptal Myofibroblasts are Responsible for Increased TLR4 Expression in a Subset of CRCs. Double-stained immunofluorescence for TLR4 (green) and vimentin (red) in normal (I), adenoma (II), and colon adenocarcinoma (III) (10x). In the stromal compartment of CRCs, immunofluorescent staining for TLR4 localized to the pericryptal myofibroblasts in a subset of samples. Image collected and cropped by CiteAb from the following publication (//www.jeccr.com/content/33/1/45), licensed under a CC-BY license.

Western Blot: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using 2 ug/mL on (A) human intestine and 6 ug/mL on (B) mouse intestine and (C) rat intestine lysate.

Flow (Intracellular): TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using PE conjugate of NBP2-27149. An intracellular stain was performed on Jurkat cells with TLR4 antibody (76B357.1) NBP2-27149PE (blue) and an isotype control MAB004 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.

Immunocytochemistry/Immunofluorescence: TLR4 Antibody (76B357.1) [NB100-56566] - RH-30 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-TLR4 Antibody (76B357.1) NB100-56566 at 1 ug/ml overnight at 4C and detected with an anti-mouse 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: TLR4 Antibody (76B357.1) [NB100-56566] - Immunofluorescent staining of TMAs. Representative tissue cores from normal (I), adenomatous polyps (II), and CRC (III and IV) are shown. Image collected and cropped by CiteAb from the following publication (//www.jeccr.com/content/33/1/45), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of rat salivary gland tissue section at 1:100 dilution. The antibody generated a membrane-cytoplasmic staining in the tissue with stronger signal in ductal epithelial cells.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Tissue section of normal human skin stained with antibody at 5 ug/mL. Membrane-cytoplasmic immunopositivity of TLR4 was primarily observed in the pigmented basel cells and the adjacent keratinocytes in the epidermal layer.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on RH30 cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human skin stained with at 5 ug/mL, peroxidase-conjugate and DAB chromogen. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of TLR4 in FFPE human colon tissue using an isotype control (top) and NB100-56566 (bottom) at 5 ug/mL.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human testis tissue stained with antibody at 5 ug/mL.

Immunohistochemistry-Frozen: TLR4 Antibody (76B357.1) [NB100-56566] - This image is TLR4(green) and nucleus(blue) at area postrema of the adult male mouse brain, x20 magnification. Primary antibody diluted 1:500. IHC-Fr image submitted by a verified customer review.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of formaldehyde fixed THP-1 cells (human monocytic leukemia cells) using 2 ug/10^6 cells TLR4 antibody (clone 76B357.1) with detection employing a donkey anti-mouse IgG (H+L) cross adsorbed secondary antibody, (DyLight 488 conjugated). Isotype control samples incubated with mouse IgG2b isotype control antibody were processed in parallal under the same assay conditions.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on Jurkat cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on Raw264.7 cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Representative images of immunohistochemical staining of pancreatic adenocarcinoma upregulated factor (PAUF) and toll-like receptor 4 (TLR4) in human ovarian cancer specimens. The top row (A,B) represents normal ovarian tissue, middle row (C,D) shows immunoreactivity of ovarian carcinomas and the bottom row (E,F) indicates immunonegativity of ovarian caricinoma. The stromal cells of normal ovarian tissue show immunonegativity for PAUF and TLR4. The scale bar represents 100 μm.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Increased toll‐like receptor 4 (TLR4) expression in the myocardium of chronic heart failure (CHF) rats. (A) TLR4 mRNA levels in infarct & remote myocardium of sham & CHF rats (n = 6/group). (B) Representative Western blot images & (C) quantification of TLR4 proteins in infarct & remote myocardium of sham & CHF rats (n = 4/group). (D) Representative immunohistochemistry images of heart sections stained with TLR4 (green) & CD45 (red). The yellow box indicates the enlarged area shown on the right (data are means ± SD, *P < 0.05, **P < 0.01 versus respective sham). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26290459), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Regulation of FOXC1 on TLR3/4 expression in mice subjected to myocardial ischaemia (MI). The siRNA against FOXC1 (si‐FOXC1) or FOXC1 adenovirus (Ad‐FOXC1) was injected into the left ventricle just after LAD ligation to generate FOXC1 knockdown (A) or FOXC1 overexpression (B), & the negative control (NC) siRNA & Ad‐GFP served as control, respectively. The mRNA (upper panel) & protein (middle & lower panel) levels of TLR3/4 were determined after 2 wk. Data are means ± SEM aP < .05, AP < .01 vs respective MI Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31517441), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Representative images of immunohistochemical staining of pancreatic adenocarcinoma upregulated factor (PAUF) & toll-like receptor 4 (TLR4) in human ovarian cancer specimens. The top row (A,B) represents normal ovarian tissue, middle row (C,D) shows immunoreactivity of ovarian carcinomas & the bottom row (E,F) indicates immunonegativity of ovarian caricinoma. The stromal cells of normal ovarian tissue show immunonegativity for PAUF & TLR4. The scale bar represents 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30111860), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Increased toll‐like receptor 4 (TLR4) expression in the surviving cardiomyocytes of chronic heart failure (CHF) rats. (A) Representative immunofluorescent images of TLR4 in cardiomyocytes isolated from sham & CHF rats. (B) TLR4 mRNA levels in cardiomyocytes isolated from sham & CHF rats. (C) Representative Western blot images & (D) quantification of TLR4 proteins in cardiomyocytes isolated from sham & CHF rats (data are means ± SD, n = 6/group, **P < 0.01 versus sham). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26290459), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Up‐regulation of FOXC1 & TLR3/4 protein levels under myocardial ischaemia. Representative Western blot images & quantitative data for FOXC1, TLR3 & TLR4 proteins in ischaemia models of mice (A), H9c2 cells (B) & NRVMs (C) are shown herein. Validation data of the antibodies for TLR3 & TLR4 are shown in Figure S1. Data are means ± SEM. n = ~4‐5/group. P values from the one‐way ANOVAs: 0.018 (TLR3 protein in mice), .002 (TLR4 protein in mice) & 0.006 (FOXC1 protein in mice). aP < .05, AP < .01 vs control Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31517441), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - (a) Cell viability of MM cells exposed to increasing doses of LPS for 24 h. (b) Immunoblotting analysis of TLR4 protein expression levels in H929 & L363 cell lines after incubation with various LPS concentrations. Probing with beta -ACTIN was used as total protein loading reference. (c) Flow cytometry analysis for TLR4 expression of MM cell lines before & after exposure to 1 μg/ml LPS for 24 h. (d) Cell viability of MM cell lines pre-treated with 0.5 μg/ml TLR4 inhibitor for 24 h before stimulation with LPS for 24 h. (e) Cell vViability of H929 & JJN3 cell lines exposed to increasing concentrations of TLR4 inhibitor for 48 h. beta -ACTIN probing was used as reference for total protein input. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30824741), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - TLR4 mRNA (a1) & protein (a2) expression in MM cell lines. (b) TLR4 mRNA expression of 16 CD138+ selected MM patients as determined by PCR & agarose gel electrophoresis. Probing with beta -ACTIN was used as total protein loading reference, whereas beta -ACTIN gene expression was used as reference for RNA input. beta -ACTIN probing & beta -ACTIN mRNA expression were used as reference for total protein & mRNA input, respectively. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30824741), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Regulation of FOXC1 on TLR expression. H9c2 cells were transfected with adenovirus or siRNA to overexpress or knock down FoxC1, with GFP adenovirus (Ad‐GFP) & negative control (NC) siRNA serving as control, respectively. A, FoxC1 overexpression increased Tlr3/4 mRNA & protein levels, under both control & ischaemic conditions. P values from the one‐way ANOVAs: <.001 (TLR3 mRNA), .002 (TLR4 mRNA), <.001 (FOXC1 mRNA), <.001 (TLR3 protein), <0.001 (TLR4 protein) & <.001 (FOXC1 protein). B, FoxC1 knockdown decreased Tlr3/4 mRNA & protein levels. P values from the one‐way ANOVAs: .001 (TLR3 mRNA), <.001 (TLR4 mRNA), <.001 (FOXC1 mRNA), .002 (TLR3 protein), <.001 (TLR4 protein) & <.001 (FOXC1 protein). C, FoxC1 overexpression up‐regulated the mRNA expression of multiple Tlr subtypes. P values from the one‐way ANOVAs: .022 (TLR1 mRNA), .047 (TLR2 mRNA), .002 (TLR3 mRNA), .013 (TLR4 mRNA), .039 (TLR5 mRNA), .003 (TLR6 mRNA), .203 (TLR7 mRNA), .078 (TLR8 mRNA) & .009 (TLR9 mRNA). Data are means ± SEM of 4 independent experiments. aP < .05, AP < .01 vs. control; bP < .05, BP < 0.01 vs. ischaemia Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31517441), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Immunofluorescent staining of TMAs. A) Low power (10x) view of NCI TMA slide stained for TLR4 (green), intestinal epithelium/pan-cytokeratin (red), & nucleus/DAPI (blue). B) Representative tissue cores from normal (I), adenomatous polyps (II), & CRC (III & IV) are shown. C & D) TLR4 staining score by tissue type & tissue compartment (stroma vs epithelium) are shown. C) TLR4 staining in the tumor stroma had a significantly higher average intensity score for stages 3 & 4 CRC when compared to stage 1. D) TLR4 staining in the tumor epithelium had a significantly higher average intensity score for stages 2 & 3 when compared to stage 1. E) TLR4 staining by compartment broken down by stage (controlling for grade) & grade (controlling for stage). Image collected & cropped by CiteAb from the following publication (//jeccr.biomedcentral.com/articles/10.1186/1756-9966-33-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Effect of Cucumis sativus L. extract (CSE) on LPS induced TLR4 expression. a Expression of TLR4 mRNA in pAECs treated with LPS (10 μg/ml) for different time (1, 7, 24 h) in the presence or absence of increasing doses of CSE (0.02; 0.2; 2 mg/ml). mRNA expression of TLR4 is determined by quantitative PCR; relative expression was calculated as fold of change in respect to the control cells & error bar represents the range of relative expression. Different letters above the bars indicate significant differences. b Representative Western Blot of TLR4 & relative housekeeping alpha -tubulin were reported. c Expression of TLR4 protein in pAECs treated with LPS (10 μg/ml) in the presence or absence of increasing doses CSE (0.02; 0.2; 2 mg/ml); data shown are representative of at least three independent experiments & represent the mean ± SEM. Different letters above the bars indicate significant differences (p < 0.05 ANOVA post hoc Tukey’s test) Image collected & cropped by CiteAb from the following publication (//bmccomplementalternmed.biomedcentral.com/articles/10.1186/s12906-018-2254-1), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Activation receptors are shed during intrinsic repair. (A) BMDM were unchallenged (control) or challenged with 500 HU/mL SLO WT, PFO or an equivalent mass of SLO ML for 10 min at 37 °C & centrifuged at 2000 × g for 5 min to yield cell pellet (C). Supernatants were spun at 100,000xg for 40 min at 4 °C to collect the high speed supernatant (S) & microvesicle pellet (M). Samples were solubilized at 95 °C in SDS-sample buffer, resolved by SDS-PAGE & transferred to nitrocellulose. Portions of the blot were probed with 6D11 anti-SLO, anti-IFN gamma R1, 4B4F12 anti-CD14, 76B357.1 anti-TLR4, O91B8 anti-MyD88, 1H4B01 anti-Trif, EPR4477 anti-Alkaline Phosphatase, MANLAC-4A7 anti-Lamin A/C, & AC-15 anti-beta -Actin antibodies followed by relevant secondary antibodies & ECL. Full-length blots are presented in Supplementary Figure S1. (B) BMDM were challenged with 500 HU/mL SLO WT or SLO N402C, SLO ML at equivalent mass to SLO WT, or SLO N402E at equivalent mass to SLO N402C for 10 min. Cell pellets (C), high speed supernatants (S) & microvesicles (M) were isolated as in (A) & probed with 6D11 anti-Streptolysin O, 76B357.1 anti-TLR4, O91B8 anti-MyD88, 1H4B01 anti-Trif, EPR4477 anti-Alkaline Phosphatase, MANLAC-4A7 anti-Lamin A/C, & AC-15 anti-beta -Actin antibodies. Full-length blots are presented in Supplementary Figure S2. The blots are representative of at least 3 independent experiments. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-24955-2), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Pericryptal Myofibroblasts are Responsible for Increased TLR4 Expression in a Subset of CRCs. A) CRCs were separated into two groups representing low- & high- stromal expression of TLR4 by IHC staining. In normal tissue, stromal TLR4 expression is mainly due to macrophages (Green: TLR4, Red: CD68, Merge: TLR4 + CD68 + DAPI (blue)). Conversely, in CRCs increased vimentin & decreased CD68 staining in the pericryptal space confirm that this signal was due to pericryptal myofibroblasts & not related to tumor-associated macrophages. B) Double-stained immunofluorescence for TLR4 (green) & vimentin (red) in normal (I), adenoma (II), & colon adenocarcinoma (III) (10×). In the stromal compartment of CRCs, immunofluorescent staining for TLR4 localized to the pericryptal myofibroblasts in a subset of samples. C) IHC staining of colon adenocarcinoma for TLR4, vimentin, & alpha -SMA (40×). Staining co-localizes to the pericryptal space, confirming the signal arises from pericryptal myofibroblasts. D, E, & F) An increase in IHC staining for alpha -SMA & vimentin was noted in CRCs when compared to normal or low grade dysplasia. A decrease in staining for CD68 positive macrophages was observed with higher degrees of dysplasia. Image collected & cropped by CiteAb from the following publication (//jeccr.biomedcentral.com/articles/10.1186/1756-9966-33-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - (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 & (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) & 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 & (e2) % proliferation of H929 & U266 cells after transfection with the pCMV6-TLR4 construct or with the empty pCMV6 vector for 48 h. beta -ACTIN probing & beta -ACTIN mRNA expression were used as reference for total protein & 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 & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30824741), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Dasatinib affects TLR4 signaling to regulate LPS-induced proinflammatory cytokine levels in BV2 microglial cells. a BV2 microglial cells were pretreated with TAK-242 (a TLR4 receptor inhibitor, 500 nM) or vehicle (1% DMSO) for 30 min, treated with dasatinib (250 nM) or vehicle (1% DMSO) for 30 min, & treated with LPS (1 μg/ml) or PBS for 5 h. The mRNA levels of COX-2 & IL-6 were analyzed by RT-PCR. b, c Quantification of the data in a (COX-2: con, n = 10; LPS, n = 10; dasatinib+LPS, n = 10; TAK-242+LPS, n = 10; TAK-242+dasatinib+LPS, n = 10; & IL-6: con, n = 14; LPS, n = 14; dasatinib+LPS, n = 14; TAK-242+LPS, n = 14; TAK-242+ dasatinib+LPS, n = 14). d BV2 microglial cells were treated with dasatinib (250 nM) or vehicle (1% DMSO) for 30 min followed by LPS (1 μg/ml) or PBS for 5.5 h, & cell surface biotinylation was conducted. e, f Quantification of the data in d (Surface TLR4: con, n = 11; LPS, n = 11; dasatinib+LPS, n = 11; Total TLR4: con, n = 16; LPS, n = 16; dasatinib+LPS, n = 16). One-way ANOVA with Tukey’s post hoc test was used to analyze significant differences. **p < 0.01, ***p < 0.001 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31655606), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - TLR4 expression in pVW-MSCs cultured with or without LPS (10 μg/ml) for 1 & 4 h & after additional 24 h of recovery after LPS removal (4hR). a: flow cytometry analysis were performed in not fixed & not permeabilized cells for TLR4 surface expression determination (TLR4: Surface) & in fixed & permeabilized cells to measure the overall TLR4 amount (TLR4: Total). Red histograms: stained cells; blue histograms: control cells. b: representative Western Blot of TLR4 & housekeeping beta -tubulin & relative quantification were presented. c: representative images of TLR4 immunostaining of pVW-MSCs cultured with or without LPS (10 μg/ml) for 4 h. pVW-MSCs nuclei were stained with Hoechst 33258 (blue). Scale bar = 10 μm. Data shown represent the mean ± SD of three biological replicates, each experiment is repeated three times. Data were analysed using one-way ANOVA followed by the Tukey’s post hoc comparison test. Different letters above the bars indicate significant differences (p < 0.05). (AU = Arbitrary Units) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31029157), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Flow Cytometry: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - TLR4 expression in pVW-MSCs cultured with or without LPS (10 μg/ml) for 1 & 4 h & after additional 24 h of recovery after LPS removal (4hR). a: flow cytometry analysis were performed in not fixed & not permeabilized cells for TLR4 surface expression determination (TLR4: Surface) & in fixed & permeabilized cells to measure the overall TLR4 amount (TLR4: Total). Red histograms: stained cells; blue histograms: control cells. b: representative Western Blot of TLR4 & housekeeping beta -tubulin & relative quantification were presented. c: representative images of TLR4 immunostaining of pVW-MSCs cultured with or without LPS (10 μg/ml) for 4 h. pVW-MSCs nuclei were stained with Hoechst 33258 (blue). Scale bar = 10 μm. Data shown represent the mean ± SD of three biological replicates, each experiment is repeated three times. Data were analysed using one-way ANOVA followed by the Tukey’s post hoc comparison test. Different letters above the bars indicate significant differences (p < 0.05). (AU = Arbitrary Units) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31029157), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - Toll‐like receptor 4 (TLR4)‐shRNA lentivirus reduced myocardial inflammation & improved heart function after myocardial infarction (MI). The rats received intra‐myocardial injection of normal saline (NS), control‐shRNA lentivirus or TLR4‐shRNA lentivirus (1 × 109 TU/ml, 100 μl/heart) just after left anterior descending coronary artery (LAD) ligation or sham operation. All examinations were performed after 4 weeks of MI. (A) Expression of green fluorescent protein (GFP; green), the marker gene carried by TLR4‐shRNA lentivirus, in the myocardium. The nuclei were counter‐stained with Hoechst 33258 (blue). (B) Representative Western blot images & quantification of TLR4 proteins in sham & chronic heart failure (CHF) myocardium. (C) tumour necrosis factor (TNF)‐ alpha & interleukin (IL)‐6 protein contents in infarct & remote myocardium. (D) Representative images of Masson's trichrome staining (upper panel) & quantification (lower panel) of post‐infarct failing hearts, showing that TLR4‐shRNA lentivirus reduced cardiac fibrosis. Cross‐sections were cut at the midhorizontal plane of the fixed paraffin‐embedded heart, & stained with Masson's trichrome reagents. (E) Infarct size of post‐infarct failing hearts. (F) Fractional shortening (%) of the left ventricle (data are means ± SD, n = 4/group, aP < 0.05, AP < 0.01 versus respective sham‐NS; BP < 0.01 versus respective CHF‐NS). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26290459), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TLR4 Antibody (76B357.1) - BSA Free [NB100-56566] - ALWPs significantly reduced LPS-induced cell-surface levels of TLR4. (A) BV2 microglial cells were pretreated with TAK-242 (a TLR4 inhibitor, 500 nM) for 30 min, followed by treatment with ALWPs (500 μg/ml) or PBS for 30 min & finally LPS (1 μg/ml) or PBS for 5 h. Total RNA was isolated, & IL-1 beta mRNA levels were measured by RT-PCR. (B) Quantification of the data from (A) (con, n = 20; LPS, n = 20; ALWPs + LPS, n = 20; TAK-242 + LPS, n = 20; TAK-242 + ALWPs + LPS, n = 20). (C) BV2 microglial cells were pretreated with ALWPs (500 μg/ml) or PBS for 30 min, followed by treatment with LPS (1 μg/ml) or PBS for 5.5 h. Cell-surface biotinylation was then conducted with a TLR4 antibody recognizing the N-terminal region of TLR4. (D) Quantification of the data from (C) (Surface TLR4: con, n = 10; LPS, n = 10; ALWPs + LPS, n = 10; Total TLR4: con, n = 4; LPS, n = 4; ALWPs + LPS, n = 4). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30319390), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ma, Pm, ChHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ma, Pm, ChHa

Applications:

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

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

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-5031A

Alt Catalog# IMG-5031A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-27149AF350
Alexa Fluor 405	NBP2-27149AF405
Alexa Fluor 488	NBP2-27149AF488
Alexa Fluor 532	NBP2-27149AF532
Alexa Fluor 594	NBP2-27149AF594
Alexa Fluor 647	NBP2-27149AF647
Alexa Fluor 700	NBP2-27149AF700
Alexa Fluor 750	NBP2-27149AF750
Allophycocyanin	NBP2-27149APC
Allophycocyanin/Cy7	NB100-56566APCCY7
Biotin	NBP2-27149B
DyLight 350	NBP2-27149UV
DyLight 405	NBP2-27149V
DyLight 488	NBP2-27149G
DyLight 550	NBP2-27149R
DyLight 594	NB100-56566DL594
DyLight 650	NBP2-27149C
DyLight 680	NBP2-27149FR
DyLight 755	NBP2-27149IR
FITC	NBP2-27149F
HRP	NBP2-27149H
Janelia Fluor 549	NB100-56566JF549
Janelia Fluor 646	NB100-56566JF646
PE	NBP2-27149PE
PE/Atto594	NB100-56566PEATT594
PE/Cy5.5	NB100-56566PECY55
PE/Cy7	NB100-56566PECY7
PerCP	NBP2-27149PCP
mFluor Violet 450 SE	NBP2-27149MFV450
mFluor Violet 500 SE	NBP2-27149MFV500
mFluor Violet 610 SE	NBP2-27149MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Azide and BSA Free	NBP2-27149
BSA Free	NB100-56566SS
Azide and BSA Free	NBP2-27149SS

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NB100-2284

CD31/PECAM-1 Antibody

(5 Reviews)

(151 Publications)

0.1 ml

NB100-2284

Staining of CD31/PECAM-1 in Rat Liver sample. IHC-P image submitted by a verified customer review.

Human lung adenocarcinoma using. Antibody at 1:100 with citrate epitope retrieval at pH 6.0.

Analysis in rat muscle blood vessels using. IHC-P image submitted by a verifed customer review.

Matrix-HA supplemented with strontium was implanted subcutaneously in mice, where immunochemistry analysis and histology occurred of the newly formed tissues. (C) Within each matrix: Matrix-HA, Matrix-8Sr-HA and Matrix-50Sr-HA, were indicated by immunostaining of CD31 for the newly formed tissues. (D) Number of vessels within each tissue was quantified using NDP view software. Unit is square mm. Immunostaining analysis of slides occurred for 2 samples per condition and 3 sections were analyzed per sample and per group of matrix. Citation: Ehret C, Aid-Launais R, Sagardoy T, Siadous R, Bareille R, Rey S, et al. (2017) Strontium-doped hydroxyapatite polysaccharide materials effect on ectopic bone formation. PLoS ONE 12(9): e0184663. //doi.org/10.1371/journal.pone.0184663

Imaging of HUVEC monolayer on glass substrate. Fixation with 4% PFA. Blocking with Goat Serum at 1:100. Incubation 2 hours at room temperature. ICC/IF image submitted by a verified customer review.

Analysis of canine kidney tissue using CD31/PECAM-1 antibody. Antibody was used at a 1:50 concentration in Casein in PBS and left at 4C overnight on paraffin embedded canine bladder tissue. HIER was performed in Tris/EDTA buffer, pH 9 for two hours at 75C. Image from verified customer review.

Analysis in pig SC cells using CD31/PECAM-1 Antibody (red). Blue color showing nucleus labeling. ICC/IF image submitted by a verified customer review.

Immunohistochemistry evaluation of CD31/PECAM-1 expression. Representative images of CD31/PECAM-1 staining of vessels in the three groups of implanted matrices (scale bar = 100 um). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-06258-0) licensed under a CC-BY license.

CD31 staining on E11.5 mouse Pharyngeal Mesoderm. Fixed with 4% PFA overnight. Blocked with 1% BSA. Primary antibody at 1:100. Secondary antibody at 1:1000, conjugated to Alexa Fluor 488. IHC-Fr image submitted by a verfied customer review.

Immunohistochemistry evaluation of CD31 expression. (a) Representative images of CD31 staining of vessels in the three groups of implanted matrices (scale bar = 100 µm). (b) Quantification of vessel density inside the implanted matrices at 5 weeks post implantation (n = 6; Average ± SD). NS and **denote Non Significant and p < 0.01, respectively.

Immunohistochemistry: Rabbit Polyclonal CD31/PECAM-1 Antibody [NB100-2284] - Staining of CD31 in human lung tissue. Nuclei in blue and CD31 (vessel) in green. Mouse anti-CD31 (1:100, overnight incubation at +4°C), followed by donkey anti-mouse 488 (1:400, 3h at RT). Image from a verified customer review.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - Antibodies selected for characterization of stromal cells & vasculature in FFPE specimens are species-specific. (A, B) Labeling of a patient colon tumor (A) & Colo205 human cell line xenograft (B) with anti-human mitochondrial antibody (single arrow- tumor cells; double arrows- stroma). (C, D) Labeling of a patient pancreatic tumor (C) & Colo205 human cell line xenograft (D) with anti-human CD34. (E, F) Labeling of a patient lung tumor (E) & Colo205 xenograft (F) with anti-human CD31. (G, H) Labeling of a patient lung tumor (G) & Colo205 xenograft (H) with anti-mouse CD34. (bars = 50 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - At the time of second passage, vessels in successfully growing xenografts of four different tumor types surveyed were of murine origin: A-C Colon; D-F Lung; G-I Pancreatic; J-L Renal Cell Carcinoma. For each tumor type, a representative section of an original patient specimen labeled with anti-huCD31 is shown (A, D, G, J). For each tumor, sections of the first passage xenografted tumor when it was resected are also shown; while no huCD31(+) vessels were identified in the xenografts (B, E, H, K), msCD34(+) vessels were abundant (C, F, I, L). (bars = 50 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - Vascularization of an engrafted patient colon tumor. A patient colon tumor was implanted in a cohort of mice & vessel development was analyzed over 8 weeks. Vessels in the original patient specimen labeled strongly for huCD31 (A) & not for msCD34 (D). Representative sections showing loss of huCD31(+) vessels (B- 4 weeks, C- 7 weeks) & presence of msCD34(+) vessels (E- 4 weeks, F- 7 weeks) are shown. The graph (G) summarizes this process; huCD31(+) vessels were rapidly lost, & by one week, mouse vessels were the predominate vessels present in the colon tumors (no data for msCD34 at 2 weeks; bars = 100 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: CD31/PECAM-1 Antibody [NB100-2284] - KCl pulse decreased membrane, but increased nuclear, detection of NHE1 in b.End3 endothelial cells.(A) Representative immunofluorescence images of bEnd.3 cells at two time-points after KCl or aCSF pulse. (B) Representative immunoblots of NHE1, alpha -tubulin, PECAM, & lamin B in cytosol, membrane, & nuclear fractions of bEnd.3 cells harvested at 5 min after KCl or aCSF pulse. (C = cytosol, M = membrane, N = nuclear) Values represent the mean ratio of NHE1 detection ± SEM (n = 11–12). (C) Representative immunoblots indicating NHE1 & alpha -tubulin as a loading control in whole cell lysate of bEnd.3 cells harvested at 5 min after KCl or aCSF pulse. Values represent the % of aCSF-treated relative expression ± SEM (n = 6). (D) Representative immunoblots of NHE1 & alpha -tubulin as a loading control in whole lysate of microvessels harvested at 90 min after cortical injection of KCl or aCSF. Values represent the % of naive relative expression ± SEM (n = 6). # denotes significantly different vs naïve (p<0.01), as assessed by one-way ANOVA (E) Intracellular pH during aCSF or KCl pulse. All data represent mean ± SEM (n = 45). (F) Extracellular pH during aCSF or KCl pulse. All data represent mean ± SEM (n = 6) in triplicate. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32469979), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: CD31/PECAM-1 Antibody [NB100-2284] - Impact of T‐AuNPs on endothelial cells in vivo. Immunofluorescent staining for endothelial cells using an anti‐CD 31 antibody revealed an intact endothelial cell monolayer in the uninjured control carotid artery (N = 3). As expected post balloon angioplasty, there is no endothelial cell monolayer of the injured left carotid arteries in the injury alone, injury + T‐AuNP, & injury + S‐AuNP treatment conditions. Green indicates autofluorescence of the elastic lamina. Magenta indicates endothelial cells. Images obtained using 25× magnification with exposure time of 400 msec. TUNEL staining for apoptosis revealed no evidence of endothelial cell apoptosis in the uninjured control carotid arteries, with a prominent endothelial cell monolayer (white arrowhead). Mild apoptosis was noted in the media of balloon‐injured left carotid arteries, as expected. Black arrows indicate apoptotic cells. Representative images obtained using 40× magnification. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28242820), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - The growth of a patient mesothelioma xenograft was supported by development of a murine vascular network. A patient mesothelioma was implanted in a cohort of mice & monitored for tumor growth. Once tumors began to actively grow, representative tumors were resected at weekly intervals & analyzed for vessel content. Staining for huCD31 was prominent in the original patient specimen (A), much reduced at 4 weeks (B) & negligible at 9 weeks (C). In contrast, patient specimens were not stained for msCD34 (D), whereas at 4 weeks large numbers of vessels stained for msCD34 (E) & by 9 weeks, msCD34 labeled vessels were predominant (F). The graph (G) summarizes the loss of detectable human vessels & acquisition of murine vessels over a 9 week period. (bars = 100 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: CD31/PECAM-1 Antibody [NB100-2284] - TGF‐ beta 1 released by MSCs contributes to induced myofibroblast differentiation & granulation tissue formationA–C2.5 × 105 of TGF‐ beta 1 siRNA or control siRNA‐transfected AT‐MSCs were intradermally injected around each of CD18−/− murine wound. PBS mock injection served as negative control. Wound tissue was harvested at day 2, 5, & 7 post‐wounding for quantification of human TGF‐ beta 1 mRNA (A) at day 2 by qPCR, total TGF‐ beta 1 (B), & active TGF‐ beta 1 (C) protein at day 5 by ELISA. Data are expressed as mean ± SEM, n = 3 wounds per group, **P < 0.01, by one‐way ANOVA with Tukey's test.D–GExpression of alpha ‐SMA (D & E) & CD31 (F & G) at days 5 & 7 by immunostaining on tissue sections. The dashed lines indicate the border of the wound bed & epidermis or eschar. e, epidermis; es, eschar; wb, wound bed. Scale bars: 100 μm. Quantification data are expressed as mean ± SEM, n = 3 wounds per group, *P < 0.05, ***P < 0.001, by one‐way ANOVA with Tukey's test.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32080965), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: Rabbit Polyclonal CD31/PECAM-1 Antibody [NB100-2284] - CD31 positive vessels in a canine splenic hemangiosarcoma.Tissue slides were loaded into automated research stainer, dewaxed and pretreated with EDTA-based epitope retrieval ER2 solution for 20 mins at 100°C. After a peroxide incubation for 5 min, the rabbit antibody against CD31 (NB100-2284) (1:100) was incubated for 60 mins at room temperature followed by Bond Polymer (anti-rabbit HRP) incubation for 8 min at room temperature. Mixed DAB reagent (Polymer Refine Detection Kit) was incubated for 10 mins, and Hematoxylin (Refine Detection Kit) counterstaining for 10 mins. After staining, sample slides were washed in water, dehydrated using ethanol gradient (70%, 90%, 100%), washed three times, and mounted in mounting medium. Image from a verified customer review.

Reactivity:

Hu, Mu, Rt, Po, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca

Applications:

, ,

Applications: , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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NBP2-22203

ERK1 Antibody (1E5)

(1 Publication)

0.1 ml

NBP2-22203

Immunohistochemistry-Paraffin: ERK1 Antibody (1E5) [NBP2-22203] - Immunohistochemical analysis of paraffin-embedded breast cancer tissues using ERK1 mouse mAb with DAB staining.

Flow Cytometry: ERK1 Antibody (1E5) [NBP2-22203] - Flow cytometric analysis of Hela cells using ERK1 mouse mAb (blue) and negative control (red).

Western Blot: ERK1 Antibody (1E5) [NBP2-22203] - Analysis using MAPK3 mAb against human ERK1 (AA: 9-143) recombinant protein. (Expected MW is 40.8 kDa)

Western Blot: ERK1 Antibody (1E5) [NBP2-22203] - Western blot analysis using ERK1 mouse mAb against Hela (1), Jurkat (2), RAW264.7 (3), HEK293 (4), K562 (5), NIH/3T3 (6), Cos7 (7) and PC-12 (8) cell lysate.

Immunohistochemistry-Paraffin: ERK1 Antibody (1E5) [NBP2-22203] - Analysis of paraffin-embedded bladder cancer tissues using ERK1 mouse mAb with DAB staining.

ELISA: ERK1 Antibody (1E5) [NBP2-22203] - Red: Control Antigen (100ng); Purple: Antigen (10ng); Green: Antigen (50ng); Blue: Antigen (100ng)

Reactivity:

Hu, Mu, Rt, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Pm

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

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NB300-144

Laminin Antibody - BSA Free

(8 Reviews)

(140 Publications)

0.1 ml

NB300-144

MxIF analysis of mouse submandibular salivary gland morphogenesis. MxIF of a developmental TMA including embryonic stages (E14, E16, E18) and postnatal stages (P5 and P20) was performed using sequential application of directly conjugated antibodies to detect multiple markers of tissue structures and cell types on the same tissue sections. Tissue compartments. The epithelium, mesenchyme, neurons, and basement membranes was detected using antibodies directed towards E-cadherin (ECAD, red), platelet-derived growth factor (PDGFR, green), beta III tubulin (bIII, magenta), and Laminin Antibody [NB300-144] (LMN, cyan), respectively. Image collected and cropped by CiteAb from the following publication (//bio.biologists.org/cgi/doi/10.1242/bio.20134309), licensed under a CC-BY license.

Staining of mouse section of cortex stained with Laminin Antibody [NB300-144] (red). Blue is DAPI staining of DNA. This antibody reveals strong staining in the basement membranes of blood vessels.

Staining of rat spinal cord and dorsal root paraformaldehyde/paraffin-embedded tissue using Laminin Antibody [NB300-144]. Pepsin antigen retrieval was performed on this tissue sample.

Analysis of rat heart cells lysates (lane 1) and 0.2 ug of purified laminin protein from mouse EHS sarcoma (lane 2) Laminin Antibody [NB300-144] recognizes 3 laminin isotypes: alpha 1 (440 kDa), beta 1 (220 kDa) and gamma 1 (220 kDa). Also recognized is a laminin binding protein at 120 kDa in both rat heart lysates and purified laminin protein. Since this protein always coexpresses with laminin this crossreactivity is irrelevant. Theoretical molecular weight of LAMA1 is 337 kDa.

IF Confocal analysis of HeLa cells using Laminin Antibody [NB300-144] (1:5). An Alexa Fluor 488-conjugated Goat to rabbit 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).

Western blot analysis of the effects of IP6, Ins, IP6 + Ins and normal saline on the levels of collagen IV, Laminin and Fibronectin, with Laminin Antibody [NB300-144]. IP6 or Ins treatment decreased the protein expression of LN and FN, and the combined IP6 + Ins treatment resulted in significantly greater effects compared with treatment with either compound alone. The Western blot membranes were stripped and reprobed for beta-actin as an internal control to confirm equal loading. (A) representative blots from one of three separate experiments; (B) relative band intensities based on densitometry. The results are expressed as the mean +/- standard deviation from three independent experiments. * p < 0.05 compared to the IP6 + Ins group; #p < 0.05 compared to the normal saline group. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2072-6643/8/5/286), licensed under a CC-BY license.

Immunofluorescence staining for extracellular matrix proteins in day 1 and day 7 HWJSC spheroids. HWJSC spheroids were cultured for either 1 day or 7 days (with 6 days of osteogenic differentiation), fixed, and stained for extracellular matrix protein: laminin (C) and counter-staining with Hoechst. Scale bars represent 100 um. Image collected and cropped by CiteAb from the following publication (journals.plos.org/plosone/article?id=10.1371/journal.pone.0184155), licensed under a CC-BY license. Using the Alexa Fluor 488 format of this antibody.

Laminin staining on an E12.5 mouse Right Ventricle. IHC-Fr image submitted by a verified customer review.

A surface stain was performed on HeLa cells with the Alexa Fluor 647 conjugate of Laminin Antibody [NB300-144AF647] (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 2.5 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Analysis of Laminin-1 expression in mouse EHS tumor crude extracts (left) and Laminin-2 expression in rat heart crude extracts (right), using Laminin Antibody [NB300-144]. The Laminin polyclonal antibody was used at 1 ug/mL. Theoretical Molecular Weight of NB300-144 is 337 kDa.

VSOP observed in perivascular-restricted spinal cord lesions with intact BBB. Immunostaining for laminin (brown) using Laminin Antibody [NB300-144] shows vascular endothelium and glia limitans of a perivascular lesion, along with infiltrating cells and VSOP (blue). Image collected and cropped by CiteAb from the following publication (//asn.sagepub.com/lookup/doi/10.1042/AN20120081), licensed under a CC-BY license.

SOX17 regulates adult muscle regeneration after injury in Pax7CreERT2/+;Sox17fl/fl mutant mice. Representative images of cryosections from regenerating adult TA muscles d28 after injury, showing immunofluorescence for PAX7+ (quiescent, arrows) cells. Scale bar, 25 Aum. Image collected and cropped by Citeab from the following publication (SOXF factors regulate murine satellite cell self-renewal and function through inhibition ofI2-catenin activity. Elife (2018)) licensed under a CC-BY license.

WNT11 regulates MMPs activities.(A–C) (Top panels): Serum-free medium was conditioned for 24 hrs by the indicated cells, concentrated 20-fold & assayed by gelatin zymography. Gelatinolytic activity is indicated by clear zones against a dark background of stained substrate. (Bottom): Whole cell extracts were immunoblotted with indicated antibodies. (A) Overexpression of Wnt11 in EMSC or BT473 cells enhances activity of MMP-9 & MMP-2. (B) Impaired activity of MMP-9 & MMP-2 in MDA-MB-231 cells (left) or EMSCs (right) stably expressing Wnt11 shRNAs & treated with DMOG. (C) WNT11 is required for MMP-9 & MMP-2 activity in MDA-MB-231 cells (left) or EMSCs (right) under normoxic & hypoxic culture conditions. (D) WNT11 regulates MMP2 protein in media. (Top): conditioned media from indicated cells & treatments. (Bottom): whole cell lysates were immunoblotted with indicated antibodies. (E) Recombinant WNT11 induces both MMP-2 protein & MMP-2 activity in media. (Top panels): Gelatin zymography & immunoblot of serum-free medium conditioned for the indicated times after recombinant WNT11 (r-WNT11) treatment. (Bottom): Whole cell lysates were immunoblotted with indicated antibodies. (F) MMP-2 inhibitor attenuated induced migration by WNT11. MDA-MB-231 cells infected with lentiviruses for stable expression of Wnt11 or GFP (n = 4) were incubated with either vehicle or 1 μM of ARP100. Media in the lower compartment had same concentration of DMSO or inhibitor. Values are mean ± s.e.m. *p < 0.05, **p < 0.01. For panels (A–D), HIF-1 alpha & HIF-2 alpha were shown as a marker of hypoxia, WNT11 normalized to alpha -Tubulin was shown. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Hypoxia induces expression of WNT11 through VHL.(A,B) Higher basal levels of WNT11 protein in Vhl-deleted cells (lenti-Cre infected Vhlf/f). EMSCs isolated from Vhlf/f mouse were infected with lentivirus carrying either GFP gene (for control) or Cre recombinase (for knockout). Non-infected cells were also used as a control. Immunoblot analysis of control or Vhl KO EMSCs treated with 0.1 mM DMOG (A), & EMSCs exposed to air (21% O2) or hypoxia (1% O2) for 24 hrs (B). Laminin, alpha -tubulin, & lamin A/C were used as loading controls, WNT11 normalized to alpha -Tubulin was shown. (C,D) Inactivation of the Vhl gene results in increased Wnt11 mRNA. Wnt11 & Vegf mRNA levels in liver (C) or duodenum (D) were measured by qPCR in Liver-VhlcKO or duodenum-VhlcKO & control mice (n = 5 per group). Values normalized to Tbp mRNA are expressed relative to tissues from control mice. For panels (C,D), values are mean ± s.e.m. *p < 0.05, **p < 0.01. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Laminin Antibody [NB300-144] - hS/PCs sequentially secrete basement membrane proteins during microstructure reorganization & function. Encapsulated hS/PCs in HA-PEGDA hydrogels initially produce laminin (red) (A–C) & collagen IV (green) (D–F) even at the single-cell state (A,D). Perlecan (green) (G–I) secretion follows to stabilize the laminin & collagen IV networks. Scalebar = 20 μm in all confocal micrographs. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31750298), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Laminin Antibody [NB300-144] - Western blot analysis of the effects of IP6, Ins, IP6 + Ins & normal saline on the levels of collagen IV, Lamininand Fibronectin. IP6 or Ins treatment decreased the protein expression of collagen IV, LN & FN, & the combined IP6 + Ins treatment resulted in significantly greater effects compared with treatment with either compound alone. The samples were probed with antibodies against p-collagen IV, p-LN, & p-FN. The Western blot membranes were stripped & reprobed for beta -actin as an internal control to confirm equal loading. (A) representative blots from one of three separate experiments; (B) relative band intensities based on densitometry. The results are expressed as the mean ± standard deviation from three independent experiments. * p < 0.05 compared to the IP6 + Ins group; #p < 0.05 compared to the normal saline group. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/2072-6643/8/5/286), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, ChHa, In, Ma, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, ChHa, In, Ma, Rb, Sh

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	NB300-144AF350
Alexa Fluor 405	NB300-144AF405
Alexa Fluor 488	NB300-144AF488
Alexa Fluor 532	NB300-144AF532
Alexa Fluor 594	NB300-144AF594
Alexa Fluor 647	NB300-144AF647
Alexa Fluor 700	NB300-144AF700
Alexa Fluor 750	NB300-144AF750
Allophycocyanin	NB300-144APC
Allophycocyanin/Cy7	NB300-144APCCY7
Biotin	NB300-144B
CoraFluor 1	NB300-144CL1
DyLight 350	NB300-144UV
DyLight 405	NB300-144V
DyLight 488	NB300-144G
DyLight 550	NB300-144R
DyLight 594	NB300-144DL594
DyLight 650	NB300-144C
DyLight 680	NB300-144FR
DyLight 755	NB300-144IR
FITC	NB300-144F
HRP	NB300-144H
Janelia Fluor 525	NB300-144JF525
Janelia Fluor 549	NB300-144JF549
Janelia Fluor 646	NB300-144JF646
Janelia Fluor 669	NB300-144JF669
PE	NB300-144PE
PE/Atto594	NB300-144PEATT594
PE/Cy5.5	NB300-144PECY55
PE/Cy7	NB300-144PECY7
PerCP	NB300-144PCP
mFluor Violet 450 SE	NB300-144MFV450
mFluor Violet 500 SE	NB300-144MFV500
mFluor Violet 610 SE	NB300-144MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB300-144SS

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

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Available Conjugates

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Marker

Modification

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Promotional Item

Species Glossary

SR-BI Antibody - BSA Free

HIF-1 alpha Antibody - BSA Free

HIF-1 alpha Antibody (2443B) - BSA Free

HIF-1 alpha Antibody (2443C) - BSA Free

Caspase-3 Antibody (31A1067) - (Pro and Active) - BSA Free

Ferroportin/SLC40A1 Antibody - BSA Free

ABCG1 Antibody - BSA Free

HIF-1 alpha Antibody (H1alpha67)

TLR4 Antibody (76B357.1) - BSA Free

CD31/PECAM-1 Antibody

ERK1 Antibody (1E5)

Laminin Antibody - BSA Free