Primary Antibodies: Novus Biologicals

AF2225

NKp46/NCR1 Antibody [Unconjugated]

(1 Review)

(75 Publications)

100 μg

AF2225

Mouse NKp46/NCR1 Antigen Affinity-purified Polyclonal Antibody induces IFN-? secretion in mouse natural killer (NK) cells activated with 25 ng/mL Recombinant Mouse IL-2 (Catalog # 402-ML) and 25 ng/mL Recombinant Mouse IL-12 (Catalog # 419-ML), in a dose-dependent manner, as measured using the Quantikine Mouse IFN-? ELISA Kit (Catalog # MIF00). The ED50 for this effect is typically 0.4-2.4 µg/mL.

NKp46/NCR1 was detected in perfusion fixed frozen sections of mouse spleen using Goat Anti-Mouse NKp46/NCR1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2225) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

<b>Genetic Strategies Validation. </b>Acutely Cultured Embryonic but Not Adult DRG Neurons Reveal Susceptibility to NK-Mediated Cytotoxicity by RAE1(A) Immunolabeling of co-culture (4 h) between embryonic (top) or adult (bottom) DRG neurons ( beta -tubulin, magenta) and either freshly isolated (control) or IL-2-stimulated natural killer (NK) cells (NKp46, green). The inset shows a high-magnification image of NK cell in contact with embryonic DRG neurite.(B) LDH-release cytotoxicity assay of acutely cultured (1 day in vitro) embryonic (top) and adult (bottom) DRG at various Effector (NK):Target (DRG) (E:T) ratios. Matched two-way ANOVA: embryonic DRG, F(1,10) = 100.01, p < 0.0001); adult DRG, F(1,10) = 1.25, p = 0.2982). Three replicate co-cultures for each DRG group.(C) Still images of in vitro time-lapse confocal Ca2+ imaging of rhodamine 3 AM-loaded embryonic (top) and adult (bottom) DRG (magenta) co-cultured with IL-2-stimulated NK cells (green) isolated from adult male NKp46-YFP mice.(D) Frequency histogram (30 s time bins) of neurite Ca2+ events in embryonic (top) and adult (bottom) DRG during NK co-culture. Cumulative area under the curve (right). Student’s paired t test; t = 2.290, p = 0.045. n = 6 fields of view from two repeat co-cultures per group.(E) RT-PCR of mRNA transcripts in freshly isolated splenic NK cells and embryonic and adult DRG.(F) qRT-PCR shows higher Raet1 mRNA expression in embryonic compared to adult DRG tissue. Student’s paired t test; t = 16.16, p < 0.0001. n = 5 mice, or replicates per group.(G) Western blot of embryonic and adult mouse DRG tissue (40 μg loading) with pan-RAE1 antibody and beta -actin control. Images are representative of three independent experiments.(H) Selective siRNA knockdown reduces RAE1 protein (top) and Raet1 mRNA (bottom) expression in embryonic DRG (2 d culture). Student’s unpaired t test; t = 9.060, p = 0.0008. n = 3 mice, or replicates per group.(I) LDH-release cytotoxicity assay of negative control or Raet1-selective siRNA knockdown embryonic DRG. Three replicate co-cultures for each siRNA group. Matched two-way ANOVA F(1,10) = 133.85, p < 0.0001).See also Figure S1. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30712871), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, , , ,

Applications: , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	AF2225U-100UG
Alexa Fluor 405	AF2225V-100UG
Alexa Fluor 488	AF2225G-100UG
Alexa Fluor 532	AF2225X-100UG
Alexa Fluor 594	AF2225T-100UG
Alexa Fluor 647	AF2225R-100UG
Alexa Fluor 700	AF2225N-100UG
Alexa Fluor 750	AF2225S-100UG
Allophycocyanin	AF2225A
Biotin	BAF2225
CoraFluor 1	AF2225CL1
DyLight 350	AF2225D
DyLight 405	AF2225E
DyLight 488	AF2225K
DyLight 550	AF2225L
DyLight 594	AF2225M
DyLight 650	AF2225W
DyLight 680	AF2225Y
DyLight 755	AF2225Z
Fluorescein	FAB2225F-025
Fluorescein	FAB2225F-100
HRP	AF2225H
Janelia Fluor 525	AF2225JF525
Janelia Fluor 549	AF2225I
Janelia Fluor 646	AF2225J
Janelia Fluor 669	AF2225JF669
PerCP	AF2225C
mFluor Violet 450 SE	AF2225MFV450
mFluor Violet 500 SE	AF2225MFV500
mFluor Violet 610 SE	AF2225MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	FAB2225F-025
Carrier Free	FAB2225F-100
Carrier Free	AF2225-SP

compare

(select up to 3 products from this page to compare)

NB100-56565

Caspase-1 Antibody (14F468) - BSA Free

(73 Publications)

0.1 mg

NB100-56565

Simple Western lane view shows a specific band for Caspase 1 in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

<b>Biological Strategies Validation. </b>Activity of Casp1 in OHT-injured and normotensive control eyes. (A) Casp1 was detected by intraocular injection FLICA660-labeled substrate (green) in vivo 24 h after injury. Bright labeling (arrows) is evident in cells in the GCL and inner nuclear layer (INL) layers of the OHT-challenged retinas, a diffuse labeling of cell processes located in the IPL. Casp1 activity is diminished in Panx1-/- (Px1-/- OHT) retinas and WT retinas treated with probenecid (WT/Pbcd) at 12 h postinjury. Image collected and cropped by CiteAb from the following publication (//www.frontiersin.org/article/10.3389/fnmol.2019.00036/full), licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>P. gingivalis and its OMVs differentially induce inflammasome signaling and pyroptosis in murine macrophages. BMM were infected as before (2 h at MOI of 25:1, see Materials and Methods) with viable P. gingivalis (Pg), heat-killed-Pg (HK-Pg), OMVs, or heat-inactivated-OMVs (HI-OMVs) and the activation of inflammasome components in the lysates [or supernatants (sup) where indicated] measured after 24 h by Western blot; beta-actin serves as a loading control throughout. Western blot data are representative of at least three independent experiments. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fcimb.2017.00351/full), licensed under a CC-BY license.

LukAB is a potent activator of Caspase 1. THP1 cells were intoxicated with 50 ng/mL LukAB for 1 hour and cell lysates were analyzed by immunoblot for Caspase 1 cleavage, which indicates activation. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.ppat.1004970) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Activity of Casp1 in OHT-injured and normotensive control eyes. (B) The analysis of the Casp1 immunolabeling (red, white arrows) in normotensive (NT control) and injured (OHT) retinas at 12 h and 24 h postinjury. Yellow arrows denote Casp1 colocalization with RGCs (RBPMS +, green) as well as with other cells at 24 h post-OHT. Bar, 25 um. Image collected and cropped by CiteAb from the following publication (//www.frontiersin.org/article/10.3389/fnmol.2019.00036/full), licensed under a CC-BY license.

P. gingivalis and its OMVs differentially induce inflammasome signaling and pyroptosis in human macrophages. MDM were infected as before (2 h at MOI of 25:1, see Materials and Methods) with viable P. gingivalis (Pg), heat-killed-Pg (HK-Pg), OMVs, or heat-inactivated-OMVs (HI-OMVs) and the activation of inflammasome components in the lysates was measured after 24 h by Western blot; beta-actin serves as a loading control throughout. Western blot data are representative of at least three independent experiments. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fcimb.2017.00351/full), licensed under a CC-BY license.

Analysis of Caspase-1 using a Caspase-1 monoclonal antibody. Human HeLa (A) and mouse NIH3T3 lysate probed with Caspase-1 antibody at 0.5 ug/ml and 2 ug/ml, respectively.

Adenocarcinoma of the rectum stained with Caspase-1 antibody (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.

Detection of Caspase-1 protein in a section of normal human colon using 5 ug/ml concentration of Caspase 1 antibody (clone 14F468). Distinct cytoplasmic staining along with some nuclear positivity was observed in crypts/mucosa, and staining was found to be more intense in the absorptive columnar epithelial cells. [10X Magnification]

Normal lung from human using 5 ug/ml concentration of Caspase 1 antibody (clone 14F468). In this representative lung section, different type of cells including pseudostratified columnar epithelium of bronchiole and the simple squamous epithelium of alveoli may be seen to develop immunoreactivity for Caspase 1. [10X Magnification]

Normal skin from human using 5 ug/ml concentration of Caspase 1 antibody (clone 14F468). Strong cytoplasmic/nuclear staining developed in all the epidermal cells, blood vessels and some cells of the dermal connective tissues layer. [10X Magnification]

Detection of Caspase-1 in a section of human ovarian cancer using 5 ug/ml concentration of Caspase 1 antibody (clone 14F468).

Tissue section of human intestine using Caspase-1 antibody (clone 14F468) at 5ug/ml concentration (1:200 dilution). The primary antibody binding to Caspase 1 in cells was detected using HRP conjugated anti-Mouse secondary antibody with DAB reagent, and the sections were further counterstained with hematoxylin for labeling cellular nuclei. This Caspase 1 antibody generated a diffused but specific cytoplasmic staining in columnar epithelia cells of villi, and a few cells depicted nuclear staining also. Only a subset of connective tissue cells in lamina propria depicted positivity (cytoplasmic) for this protein.

Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - Glucose fluctuations promoted the activation of NLRP3 inflammasome. (A) Immunofluorescence staining of NLRP3-ASC interaction in rat hearts. The heart sections were labeled with anti-NLRP3 (red), anti-ASC (green), & DAPI (blue) (n = 3 per group). (B–D) The mRNA levels of NLRP3, IL-1 beta & IL-18 in rat hearts of the three groups (n = 4 per group). (E–H) The protein expressions of NLRP3, ASC & cleaved caspase-1 in rat hearts of the three groups (n = 4 per group). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35592403), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - Caspase-1 expression increases with age in human fibroblasts: a Immunoblot analysis of caspase-1 & ASC in human fibroblasts obtained from a donor at the ages of 49, 52 & 64 y/o. b Immunocytochemistry image of human fibroblasts stained for caspase-1 (green) & Phalloidin (red) from a donor at three different ages (49, 52 & 64 y/o). Bar graph: 100 μm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30473634), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - Aging alters expression of NLRP1 inflammasome components. Representative immunoblots & densitometric analysis of immunoblots for caspase-1 (Casp1), caspase-11 (Casp11), NLRP1, ASC, cleaved XIAP, P2X7 receptor (P2X7R) & pannexin-1 (PanX1) in brain lysates of young (Y) & aged (A) animals. Protein levels of cleaved caspase-1 & -11 are higher in aged animals compared to young. Protein levels of NLRP1 & ASC did not change with age whereas P2X7 receptor, the pannexin-1 protein & the cleaved fragment of XIAP are higher in aged animals than in young animals. beta -Tubulin was used as an internal standard & control for protein loading. Data are presented as mean +/- SEM. *p < 0.05, **p < 0.01, ***p < 0.005 compared with young. n = 6 per group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22133203), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - Role of NF-kappa B in glucose fluctuation-induced inflammasome-related myocardial fibrosis. (A) Immunofluorescence staining of NF-kappa B/p65 nucleation (n = 3 per group). (B,C) The phosphorylation level of NF-kappa B/p65 in rat hearts of the three groups (n = 4 per group). (D) Immunofluorescence staining of intracellular NLRP3–ASC interaction in the GF & GF + TPCA-1 groups. The NRCFs were labeled with anti-NLRP3 (red), anti-ASC (green), & DAPI (blue) (n = 3 per group). (E–G) After application of TPCA-1 (0.5 μM), the protein expressions of cleaved caspase-1 & collagen I were measured by western blot (n = 3 per group). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35592403), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - Inflammasome proteins are elevated in the cytosolic fraction of aged mice: a Representative image of immunoblot analyses of inflammasome proteins in the cytosolic fraction of the cortex of young (Y) & aged (A) mice. Quantification of immunoblot analysis of NLRC4 (b), caspase-1 (c), ASC (d), IL-18 (e) in the cortex. a Representative image of immunoblot analyses of inflammasome proteins in the cytosolic fraction of the hippocampus of young (Y) & aged (A) mice. Quantification of immunoblot analysis of NLRC4 (g), caspase-1 (h), caspase-11 (i), ASC (j), IL-1 beta (k) in the hippocampus. Data presented as mean+/-SEM. N = 5 per group. *p < 0.05 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30473634), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - NLRP3 inhibition with MCC950 reversed myocardial fibrosis induced by glucose fluctuations. (A) Representative images of hematoxylin & eosin (HE) staining for myocardial tissue in the FBG + NaCl & FBG + MCC950 groups (n = 3 per group). (B–C) Representative images of myocardial fibrosis & fibrosis area (%) in the FBG + NaCl & FBG + MCC950 groups (n = 4 per group). (D–K) The protein expressions of NLRP3, cleaved caspase-1, TGF-beta 1, collagen I & collagen III in the FBG + NaCl & FBG + MCC950 groups (n = 6 per group). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35592403), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - Inflammasome proteins are elevated in the cytosolic fraction of aged mice: a Representative image of immunoblot analyses of inflammasome proteins in the cytosolic fraction of the cortex of young (Y) & aged (A) mice. Quantification of immunoblot analysis of NLRC4 (b), caspase-1 (c), ASC (d), IL-18 (e) in the cortex. a Representative image of immunoblot analyses of inflammasome proteins in the cytosolic fraction of the hippocampus of young (Y) & aged (A) mice. Quantification of immunoblot analysis of NLRC4 (g), caspase-1 (h), caspase-11 (i), ASC (j), IL-1 beta (k) in the hippocampus. Data presented as mean+/-SEM. N = 5 per group. *p < 0.05 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30473634), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - Role of inflammasome in glucose fluctuation-induced myocardial fibrosis. (A) Immunofluorescence staining of intracellular NLRP3–ASC interaction examined by confocal microscopy. The NRCFs were labeled with anti-NLRP3 (red), anti-ASC (green), & DAPI (blue) (n = 3 per group). (B–G) After siRNA targeting NLRP3 transfection, the protein expressions of NLRP3, cleaved caspase-1 & collagen I were measured by western blot (n = 3 per group). (H) Representative images of immunofluorescence staining of intracellular alpha -SMA in the NG, HG, GF, & GF + siRNA groups (n = 3 per group). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35592403), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - Epirubicin suppresses the NLRP3 inflammasome in presence of autophagy inhibitors. (A) LPS/ATP-induced IL-1 beta secretion from PM in the absence or presence of bafilomycin & 3-MA is shown. Means ± SEM from 10 (IL-1 beta ) independent experiments are shown. (B) Representative immunoblot of LC3b after epirubicin & LPS/ATP treatments. (C,D) Representative immunoblots of inflammasome & autophagy components after inhibition with (C) bafilomycin or (D) 3-MA in PM whole cell lysates. Cells were treated with indicated concentrations of epirubicin treatment over 24 h, followed by LPS (10 ng/mL) in the absence or presence of bafilomycin (300 nM) or 3-MA (10 mM) for 3 h & NLRP3 activation by ATP (1 mM, 1 h). Whole cell lysates were subjected to SDS-PAGE (10% gel) & processed for immunoblot analysis with specific antibodies. Here, beta -actin served as loading control. Statistical tests: one-sample t-test (A) ** p < 0.01, *** p < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31905600), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - Genetic or pharmacologic disruption of Caspase 1 blocks LukAB-induced cytokine secretion but not cell death.(A) THP1 cells were transfected with siRNA against Caspase 1, ASC or a non-targeting sequence. Cell lysates were collected & analyzed by immunoblot to confirm knockdown of pro-Caspase 1 & ASC. (B) THP1 siRNA cells were incubated with propidium iodide & intoxicated with 50 ng/mL LukAB for 1 hour then analyzed by flow cytometry. (C & D) THP1 siRNA cells were either primed with LTA (500ng/mL) (C) or untreated (D) then intoxicated with LukAB (50ng/mL) for 1 hr before culture supernatants were analyzed for release of IL-1 beta (C) or IL-18 (D). (E & F) Primary CD14+ human monocytes were incubated with the indicated concentration of z-YVAD-FMK or VX-765 for 30 minutes. Primary monocytes were incubated in the presence of propidium iodide (E) then intoxicated with LukAB (50ng/mL) & analyzed by flow cytometry. (F) Primary monocytes were intoxicated with LukAB (50ng/mL) & culture supernatants were analyzed for release of IL-18. Propidium iodide staining & IL-18 secretion are reported as a fraction of measurement in primary cells not treated with inhibitor. Bars represent the mean ± standard error of the mean for at least two independent experiments, each performed in triplicate. Asterisks indicate significance at a p-value of ≤ 0.05 by Tukey’s multiple comparisons post-test for 1-way or 2-way ANOVA, as appropriate. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1004970), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: Caspase-1 Antibody (14F468) - BSA Free [NB100-56565] - Probenecid reduces protein expression of NLRP1 inflammasome & ameliorates spatial learning deficits in aged rats. (A) Representative immunoblots of cleaved caspase-1, pannexin1 & P2X7R in hippocampal lysates of vehicle (Veh)-treated & probenecid (Pr)-treated 18-month-old rats. beta -tubulin was used as an internal control. (B) Densitometric analysis of immunoblots from brain lysates of cleaved caspase-1 (Casp1), P2X7 receptor (P2X7R), & pannexin1 (PanX1). (C-D) Aged animals underwent behavioral testing following either probenecid or vehicle treatment. (C) In a hippocampal-dependent spatial learning task via Morris water maze, latency to platform was measured on days 1-3 & 8-10. Probenecid-treatment improved latency to platform measured on the final day of testing (D) Mean path length was determined on day 10 of testing & probenecid-treated rats demonstrated significantly shorter mean path lengths than vehicle-treated controls. Drug treatment was administered twice daily for 3 days (days 7-9). Data are presented as mean +/- SEM *p < 0.05, **p < 0.005 compared to vehicle. N = 6-8/per group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22133203), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , ,

Applications: , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-5028

Alt Catalog# IMG-5028

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-56565AF350
Alexa Fluor 405	NB100-56565AF405
Alexa Fluor 488	NB100-56565AF488
Alexa Fluor 532	NB100-56565AF532
Alexa Fluor 594	NB100-56565AF594
Alexa Fluor 647	NB100-56565AF647
Alexa Fluor 700	NB100-56565AF700
Alexa Fluor 750	NB100-56565AF750
Allophycocyanin	NB100-56565APC
Biotin	NB100-56565B
CoraFluor 1	NB100-56565CL1
DyLight 350	NB100-56565UV
DyLight 405	NB100-56565V
DyLight 488	NB100-56565G
DyLight 550	NB100-56565R
DyLight 594	NB100-56565DL594
DyLight 650	NB100-56565C
DyLight 680	NB100-56565FR
DyLight 755	NB100-56565IR
FITC	NB100-56565F
HRP	NB100-56565H
Janelia Fluor 525	NB100-56565JF525
Janelia Fluor 549	NB100-56565JF549
Janelia Fluor 646	NB100-56565JF646
Janelia Fluor 669	NB100-56565JF669
PE	NB100-56565PE
PerCP	NB100-56565PCP
mFluor Violet 450 SE	NB100-56565MFV450
mFluor Violet 500 SE	NB100-56565MFV500
mFluor Violet 610 SE	NB100-56565MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-56565SS
Azide and BSA Free	NBP2-33230

compare

(select up to 3 products from this page to compare)

AF2419

PDX-1/IPF1 Antibody [Unconjugated]

(75 Publications)

100 μg

AF2419

Western blot shows lysates of beta TC-6 mouse beta cell insulinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human PDX-1/IPF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2419) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for PDX-1/IPF1 at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Simple Western lane view shows lysates of beta TC-6 mouse beta cell insulinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for PDX-1/IPF1 at approximately 48 kDa (as indicated) using 10 µg/mL of Goat Anti-Human PDX-1/IPF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2419) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

PDX-1/IPF1 was detected in immersion fixed BG01V human embryonic stem cells differentiated into pancreatic progenitor cells using Goat Anti-Human PDX-1/IPF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2419) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Goat IgG Secondary Antibody (green; Catalog # NL003) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

PDX-1/IPF1 was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer tissue using Goat Anti-Human PDX-1/IPF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2419) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei in cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Differentiation of MODY3-iPSCs into pancreatic beta cells.(A) scheme of 6 step differentiation protocol (B), expression pattern of HNF1A mRNA during differentiation process, (C) Phase-contrast image of spheroids of differentiated MODY3-iPS-beta cells, (D, E) Immunocytochemistry of differentiated MODY3-iPS-beta cells for pancreatic beta cell markers (D) C-peptide, PDX1, NKX6.1 and DAPI staining, (E) HNF1A and DAPI staining. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31145732), licensed under a CC-BY license. Not internally tested by R&D Systems.

Formalin-fixed paraffin-embedded tissue sections of human pancreas were probed for PDX-1 mRNA (ACD RNAScope Probe, catalog #437088; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using goat anti-human PDX-1 polyclonal antibody (R&D Systems catalog # AF2419) at 1ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to islet cells and exocrine glands.

GP2 and CD142 comparative profiles. a–j Flow cytometry analyses of H1-derived day 13 PP cultures obtained using 10 mM nicotinamide (a–e, optimal differentiation) and 3.3 mM nicotinamide (f–j, suboptimal differentiation) during stage 4 of differentiation. Cells were stained with anti-GP2 and anti-CD142 and then fixed and stained for the intracellular markers PDX1 and NKX6-1. IgG controls are shown on the left of each panel. Green boxes highlight the low percentage of GP2+/PDX− cells (c–h), red box highlights the presence of CD142+PDX1− cells (j). Abbreviations: GP2, pancreatic secretory granule membrane major glycoprotein 2; CD142, tissue factor; NKX6-1, NK6 homeobox 1; PDX1, pancreatic, and duodenal homeobox 1 Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28835709), licensed under a CC-BY license. Not internally tested by R&D Systems.

(A) Differentiation markers for the endoderm (FOXA2, PDX1), mesoderm ( alpha -actin, brachyury), and ectoderm ( beta 3 Tubulin, Otx2) were detected for the MUCG01, MUCG02, and MUCG03 clinical-grade hESC lines by immunocytochemistry. (B) Standard karyotypes without chromosomal aberration were detected for all hESC lines MUCG01: 46,XX; MUCG02: 46,XX; and MUCG03: 46,XY. (C) Pluripotency markers Oct3/4, Sox2, and Nanog were detected for the MUCG01, MUCG02, and MUCG03 clinical-grade hESC lines by immunocytochemistry. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36293356), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, , , ,

Applications: , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	AF2419U-100UG
Alexa Fluor 405	AF2419V-100UG
Alexa Fluor 488	AF2419G-100UG
Alexa Fluor 532	AF2419X-100UG
Alexa Fluor 594	AF2419T-100UG
Alexa Fluor 647	AF2419R-100UG
Alexa Fluor 700	AF2419N-100UG
Alexa Fluor 750	AF2419S-100UG
Allophycocyanin	AF2419A
Biotin	BAF2419
CoraFluor 1	AF2419CL1
DyLight 350	AF2419D
DyLight 405	AF2419E
DyLight 488	AF2419K
DyLight 550	AF2419L
DyLight 594	AF2419M
DyLight 650	AF2419W
DyLight 680	AF2419Y
DyLight 755	AF2419Z
FITC	AF2419F
HRP	AF2419H
Janelia Fluor 525	AF2419JF525
Janelia Fluor 549	AF2419I
Janelia Fluor 646	AF2419J
Janelia Fluor 669	AF2419JF669
PE	AF2419P
PerCP	AF2419C
mFluor Violet 450 SE	AF2419MFV450
mFluor Violet 500 SE	AF2419MFV500
mFluor Violet 610 SE	AF2419MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF2419-SP

compare

(select up to 3 products from this page to compare)

AF594

IL-21 Antibody [Unconjugated]

(81 Publications)

100 μg

AF594

IL‑21 was detected in immersion fixed concanavalin A-stimulated mouse splenocytes using 15 µg/mL Mouse IL‑21 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF594) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained (green). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Recombinant Mouse IL-21 (Catalog # 594-ML) stimulates proliferation in the NK-92 human natural killer lymphoma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Mouse IL-21 (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse IL-21 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF594). The ND50 is typically 0.2-2 µg/mL.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, , ,

Applications: , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	AF594U-100UG
Alexa Fluor 405	AF594V-100UG
Alexa Fluor 488	AF594G-100UG
Alexa Fluor 532	AF594X-100UG
Alexa Fluor 594	AF594T-100UG
Alexa Fluor 647	AF594R-100UG
Alexa Fluor 700	AF594N-100UG
Alexa Fluor 750	AF594S-100UG
Allophycocyanin	AF594A
Biotin	BAF594
CoraFluor 1	AF594CL1
DyLight 350	AF594D
DyLight 405	AF594E
DyLight 488	AF594K
DyLight 550	AF594L
DyLight 594	AF594M
DyLight 650	AF594W
DyLight 680	AF594Y
DyLight 755	AF594Z
FITC	AF594F
HRP	AF594H
Janelia Fluor 525	AF594JF525
Janelia Fluor 549	AF594I
Janelia Fluor 646	AF594J
Janelia Fluor 669	AF594JF669
PE	AF594P
PerCP	AF594C
mFluor Violet 450 SE	AF594MFV450
mFluor Violet 500 SE	AF594MFV500
mFluor Violet 610 SE	AF594MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF594-SP

compare

(select up to 3 products from this page to compare)

AF1019

PD-L1 Antibody [Unconjugated]

(3 Reviews)

(70 Publications)

100 μg

AF1019

Western blot shows lysates of RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 10 µg/mL LPS for 4 hours. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Mouse PD-L1/B7-H1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1019) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for PD-L1/B7-H1 at approximately 50-55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

<b>Biological Strategies Validation. </b>RAW 264.7 mouse monocyte/macrophage cell line either treated with LPS overnight (filled histogram) or untreated (open histogram) was stained with Goat Anti-Mouse PD-L1/B7-H1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1019), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). View our protocol for Staining Membrane-associated Proteins.

HEK293 human embryonic kidney cell line transfected with either (A) mouse PD-L1/B7-H1 or (B) irrelevant transfectants and eGFP was stained with Goat Anti-Mouse PD-L1/B7-H1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1019) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). Quadrant markers were set based on control antibody staining (Catalog # AB-108-C). View our protocol for Staining Membrane-associated Proteins.

<b>Biological Strategies Validation. </b>Pyruvate kinase isoform M2 (PKM2) is required for LPS-induced expression of PD-L1. RAW cells (0.5 × 106 cells/ml) (A), bone marrow-derived macrophages (BMDMs) (B), or bone marrow-derived dendritic cells (C) were treated with TEPP-46 at 50 µM for 1 h prior to stimulation with LPS (100 ng/ml, 24 h), lysed and assayed for expression of PD-L1 by flow cytometry [(A–C) top panels] and pdl1 mRNA by RT-PCR [(A,B) bottom panels]. Statistical analysis was performed on cells from three separate experiments. Error bars represent mean ± SEM. BMDM cells were transfected with scrambled control (SC) or anti-PKM2 siRNA (D). 24 h post-transfection, cells were stimulated with LPS (100 ng/ml) for 24 h after which expression of PKM2 (top panel) and PDL1 (middle panel) was measured by western blot. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29081778), licensed under a CC-BY license. Not internally tested by R&D Systems.

Formalin-fixed paraffin-embedded tissue sections of mouse intestines were probed for PD-L1 mRNA (ACD RNAScope Probe, catalog #420508; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using goat anti-mouse PD-L1 polyclonal antibody (R&D Systems catalog # AF1019) at 3ug/mL with one-hour incubation at room temperature followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to lamina propria of villi.

Protumor roles for TAMs and granulocytic myeloid-derived suppressor cells are obviated in neoantigen+ PDA.(A) Immunofluorescence staining of CB+ (nAg+) or CB– (nAg–) orthotopic tumors on day 21. Arrows show CD8+ T cell contact with a macrophage. Scale bar: 50 μm. Original magnification of insets, 2.25×. The number of CD8+ T cell and TAM (F4/80+) contacts per field of view. The box plots depict the minimum and maximum values (whiskers), the upper and lower quartiles, and the median. The length of the box represents the interquartile range. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35393950), licensed under a CC-BY license. Not internally tested by R&D Systems.

Representative images (×20 magnification) of H&E and PD-L1 IHC-stained sections (A-C): (A) Nude mouse lymph node showing normal histology (H&E) and diffuse staining for PD-L1 (mouse) with vessels and cells within the paracortical regions displaying increased staining intensity; (B) Nude mouse spleen showing normal histology (H&E) and intense membranous staining for PD-L1 (mouse) with the most intense staining at the periphery of the white pulp; (C) MDA-MB231 tumor showing a viable region (H&E necrotic regions were observed in all sections) with membranous and cytoplasmic staining for PD-L1 (human) on the tumor cells; (D) IHC quantitative analysis (staining intensity score, H-score) of PD-L1 expression levels in lymph nodes, spleen, and MDA-MB231 tumors from mouse xenografts; each bar represents the mean H-score ± standard deviation (SD; n = 3, spleen and lymph nodes; n= 6, tumors). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31044647), licensed under a CC-BY license. Not internally tested by R&D Systems.

Human pancreas tissue from type 1 diabetic subjects express PD-L1. Human pancreas sections from type 1 diabetic (T1D), autoantibody positive (AA+), type 2 diabetic (T2D), and non-diabetic controls (NDB) were obtained from the Network for Pancreatic Organ Donation (nPOD) and stained for T cell markers (CD4, CD8), insulin, and PD-L1. Shown are representative islets from each group with 7–15 unique islets analyzed from three independent experiments with one patient per group. Scale bar corresponds to 20 µm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29844327), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, , , ,

Applications: , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	AF1019U-100UG
Alexa Fluor 405	AF1019V-100UG
Alexa Fluor 488	AF1019G-100UG
Alexa Fluor 532	AF1019X-100UG
Alexa Fluor 594	AF1019T-100UG
Alexa Fluor 647	AF1019R-100UG
Alexa Fluor 700	AF1019N-100UG
Alexa Fluor 750	AF1019S-100UG
Allophycocyanin	FAB1019A-025
Allophycocyanin	FAB1019A-100
Biotin	BAF1019
CoraFluor 1	AF1019CL1
DyLight 350	AF1019D
DyLight 405	AF1019E
DyLight 488	AF1019K
DyLight 550	AF1019L
DyLight 594	AF1019M
DyLight 650	AF1019W
DyLight 680	AF1019Y
DyLight 755	AF1019Z
HRP	AF1019H
Janelia Fluor 525	AF1019JF525
Janelia Fluor 549	AF1019I
Janelia Fluor 646	AF1019J
Janelia Fluor 669	AF1019JF669
PE	AF1019P
PerCP	AF1019C
mFluor Violet 450 SE	AF1019MFV450
mFluor Violet 500 SE	AF1019MFV500
mFluor Violet 610 SE	AF1019MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF1019-SP
Carrier Free	FAB1019A-100
Carrier Free	FAB1019A-025

compare

(select up to 3 products from this page to compare)

NBP3-07012

Noxa Antibody (114C307.1R) - BSA Free

(73 Publications)

0.1 mg

NBP3-07012

Flow Cytometry: Noxa Antibody (114C307.1R) [NBP3-07012] - An intracellular stain was performed on Ntera2 cells with Noxa Antibody [114C307.1R] NBP3-07012 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Immunocytochemistry/Immunofluorescence: Noxa Antibody (114C307.1R) [NBP3-07012] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Noxa Antibody (114C307.1R) NBP3-07012 at 2 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-Paraffin: Noxa Antibody (114C307.1R) - BSA Free [NBP3-07012] - Immunohistochemical analysis of bouin-fixed, paraffin-embedded chronic lymphocytic leukemia (CLL) xenograft stained with Noxa antibody (1:2000), peroxidase-conjugate and DAB chromogen. Staining for 2 hr at RT was used.

Western Blot: Noxa Antibody (114C307.1R) - BSA Free [NBP3-07012] - Detection by Western blot in RL-7 cells (a follicular lymphoma).

Flow Cytometry: Noxa Antibody (114C307.1R) - BSA Free [NBP3-07012] - Flow cytometry after an intracellular stain was performed on MCF7 cells with Noxa [114C307.1] antibody NB600-1159AF700 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to Alexa Fluor 700.

Flow Cytometry: Noxa Antibody (114C307.1R) - BSA Free [NBP3-07012] - Flow cytometry after an intracellular stain was performed on HeLa cells with Noxa [114C307.1] antibody NB600-1159AF700 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to Alexa Fluor 700.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-1159AF350
Alexa Fluor 405	NB600-1159AF405
Alexa Fluor 488	NB600-1159AF488
Alexa Fluor 532	NB600-1159AF532
Alexa Fluor 594	NB600-1159AF594
Alexa Fluor 647	NB600-1159AF647
Alexa Fluor 700	NB600-1159AF700
Alexa Fluor 750	NB600-1159AF750
Allophycocyanin	NB600-1159APC
Biotin	NB600-1159B
Biotin	NBP2-24790
CoraFluor 1	NB600-1159CL1
DyLight 350	NB600-1159UV
DyLight 405	NB600-1159V
DyLight 488	NB600-1159G
DyLight 550	NB600-1159R
DyLight 594	NB600-1159DL594
DyLight 650	NB600-1159C
DyLight 680	NB600-1159FR
DyLight 755	NB600-1159IR
FITC	NB600-1159F
HRP	NB600-1159H
Janelia Fluor 525	NB600-1159JF525
Janelia Fluor 549	NB600-1159JF549
Janelia Fluor 646	NB600-1159JF646
Janelia Fluor 669	NB600-1159JF669
PE	NB600-1159PE
PerCP	NB600-1159PCP
mFluor Violet 450 SE	NB600-1159MFV450
mFluor Violet 500 SE	NB600-1159MFV500
mFluor Violet 610 SE	NB600-1159MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Non-Recombinant Monoclonal	NB600-1159SS
Azide and BSA Free	NBP2-33250
Non-Recombinant Monoclonal	NB600-1159
BSA Free	NBP3-07012SS

compare

(select up to 3 products from this page to compare)

NB600-582

SREBP1 Antibody (2A4) - BSA Free

(73 Publications)

0.1 ml

NB600-582

Simple Western: SREBP1 Antibody (2A4) [NB600-582] - Lane view shows a specific band for SREBP1 in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

<b>Genetic Strategies Validation. </b>Western Blot: SREBP1 Antibody (2A4) [NB600-582] - KO and OE of SREBP1 influenced lipid metabolism in Bel-7402 cells. The protein level of lipid-associated genes in SREBP1-KO, SREBP1-OE, and WT Bel-7402 cells. Data were expressed as mean -/+ standard error of the mean. *P < 0.05, **P < 0.01, and ***P < 0.001 vs. WT group, respectively.

Western Blot: SREBP1 Antibody (2A4) - BSA Free [NB600-582] - Effect of GEN on the expression of Nrf2‐related signalling proteins in OA (660 μmol/L) & PA (330 μmol/L)‐induced HepG2 cells. GEN was added into HepG2 cells prior 1 h to stimulation of OA & PA for 18 h. A‐C, Protein expression of Nrf2, PPAR alpha , PPAR gamma in nucleus & HO‐1 in cytoplasm was detected by Western blot. (D & E) Protein expression of P‐ACC, ACC, P‐AKT, AKT, P‐AMPK alpha , AMPK alpha , P‐AMPK beta & AMPK beta was detected by Western blot. F‐H, Protein expression of PI3K, P‐mTORC, mTORC, P‐S6K, S6K, P‐S6, S6, SREBP‐1c & HMGB1 was detected by Western blot. The similar results were collected from three dependent experiments. All data were expressed by mean ± SEM (n = 5 in each group). ##P < .01 vs Control Group; *P < .05 & **P < .01 vs OA & PA Group Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32293113), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SREBP1 Antibody (2A4) - BSA Free [NB600-582] - KO & OE of SREBP1 influenced lipid metabolism in Bel-7402 cells. (A) Total cellular TG content of SREBP1-KO, SREBP1-OE, & WT Bel-7402 cells. (B) HPLC-MS determination of the SFAs to MUFAs ratios (palmitic acid to palmitoleic acid, 16:0 to 16:1) (stearic acid to oleic acid, 18:0 to 18:1) as well as the content of 16:0, 16:1, 18:0, & 18:1 in SREBP1-KO, SREBP1-OE, & WT Bel-7402 cells. (C) The transcript level of lipid-associated genes in SREBP1-KO, SREBP1-OE, & WT Bel-7402 cells. (D & E) The protein level of lipid-associated genes in SREBP1-KO, SREBP1-OE, & WT Bel-7402 cells. Data were expressed as mean ± standard error of the mean. *P < 0.05, **P < 0.01, & ***P < 0.001 vs. WT group, respectively. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31297058), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SREBP1 Antibody (2A4) - BSA Free [NB600-582] - Construction of knockout (KO) & overexpression (OE) cell lines. (A) The sequencing results of KO positive clone cells; (B) The messenger RNA (mRNA) level of SREBP1 in wild type (WT) & OE Bel-7402 cells. Data were expressed as mean ± standard error of the mean. ***P < 0.001 vs. WT group; (C & D) The protein level of SREBP1 in WT, OE, & KO Bel-7402 cells. Data were expressed as mean ± standard error of the mean. ***P < 0.001 vs. WT group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31297058), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SREBP1 Antibody (2A4) - BSA Free [NB600-582] - p75NTR stimulation activated SREBP1 & SREBP2 in Huh7 hepatocyte cells.Human Huh7 cells were stimulated with 10 ng/ml pro-NGF or 50 ng/ml NGF for 16 h followed by immunoblotting using antibodies to detect the presence of cleaved/activated SREBP1 & SREBP2. a SREBP1, immunoblots & b quantification. Values are mean ± SD, n = 3. **P < 0.01 & *p < 0.05 for treated vs. control cells. c, d SREBP2. Cells were treated with 10 ng/ml pro-NGF in the absence or presence of 0.5 μM PF429242 (PF), which is a selective inhibitor of the Site 1 protease (S1P). c Immunoblots & d quantification. PF decreased the amount of processed SREBP2 in these cells; however, addition of pro-NGF increased the relative level of cleaved SREBP2. Values are mean ± SD, n = 3. *p < 0.05 for treated vs. the corresponding control Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31296846), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SREBP1 Antibody (2A4) - BSA Free [NB600-582] - Effect of GEN on the protein expression of Nrf2, ACC & mTORC signalling pathways in tyloxapol‐induced mice. GEN (50, 75, 100 mg/kg) was administered to WT or Nrf2−/− mice prior 1 h to stimulation of tyloxapol (500 mg/kg) for 18 h. A‐C, Protein expression of Nrf2, PPAR alpha , PPAR gamma in nucleus & HO‐1 in cytoplasm of WT mice was detected by Western blot. (D & E) Protein expression of Nrf2, PPAR alpha & PPAR gamma in Nrf2−/− mice was detected by Western blot. F‐H, Protein expression of P‐ACC, ACC, P‐AKT, AKT, P‐AMPK alpha , AMPK alpha , P‐AMPK beta , AMPK beta in WT & Nrf2−/− mice was detected by Western blot. I‐M, Protein expression of PI3K, P‐mTORC, mTORC, P‐S6K, S6K, P‐S6, S6, SREBP‐1c & HMGB1 WT & Nrf2−/− mice was detected by Western blot. The similar results were collected from three dependent experiments. All data were expressed by mean ± SEM (n = 5 in each group). ##P < .01 vs Control group; *P < .05 & **P < .01 vs Tyloxapol group Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32293113), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ca, Ch, Ha, Pm, SyHa

Species Glossary

Reactivity: Hu, Mu, Rt, Ca, Ch, Ha, Pm, SyHa

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	NB600-582AF350
Alexa Fluor 405	NB600-582AF405
Alexa Fluor 488	NB600-582AF488
Alexa Fluor 532	NB600-582AF532
Alexa Fluor 594	NB600-582AF594
Alexa Fluor 647	NB600-582AF647
Alexa Fluor 700	NB600-582AF700
Alexa Fluor 750	NB600-582AF750
Allophycocyanin	NB600-582APC
Biotin	NB600-582B
CoraFluor 1	NB600-582CL1
DyLight 350	NB600-582UV
DyLight 405	NB600-582V
DyLight 488	NB600-582G
DyLight 550	NB600-582R
DyLight 594	NB600-582DL594
DyLight 650	NB600-582C
DyLight 680	NB600-582FR
DyLight 755	NB600-582IR
FITC	NB600-582F
HRP	NB600-582H
Janelia Fluor 525	NB600-582JF525
Janelia Fluor 549	NB600-582JF549
Janelia Fluor 646	NB600-582JF646
Janelia Fluor 669	NB600-582JF669
PE	NB600-582PE
PerCP	NB600-582PCP
mFluor Violet 450 SE	NB600-582MFV450
mFluor Violet 500 SE	NB600-582MFV500
mFluor Violet 610 SE	NB600-582MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-582SS
Azide and BSA Free	NBP2-80959

compare

(select up to 3 products from this page to compare)

NB100-39002

FoxP3 Antibody - BSA Free

(2 Reviews)

(66 Publications)

0.1 ml

NB100-39002

Immunohistochemistry: FoxP3 Antibody [NB100-39002] - CCR10-expressing CD4+CD25+FOXP3+ Treg cells are enriched in the spinal cord after SCI. Double immunostaining of FOXP3 (red) and CCR10 (green) in the spinal cord sections after sham or SCI surgery. The merged images are also shown. Arrows indicate cells with positive staining and colocalization. Scale bar, 100 i1/4m. Data are mean A+- SD. Image collected and cropped by Citeab from the following publication (CCL28 promotes locomotor recovery after spinal cord injury via recruiting regulatory T cells. Aging (Albany NY) (2019) licensed under a CC-BY license.

Flow Cytometry: FoxP3 Antibody [NB100-39002] - An intracellular stain was performed on SK-MEL-28 cells with NB100-39002 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody.

Immunohistochemistry-Paraffin: FoxP3 Antibody [NB100-39002] - Tissue section of human tonsil using 1:200 dilution of rabbit anti-FoxP3 antibody. The staining was developed with HRP-labeled anti-rabbit IgG secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin. This FoxP3 antibody generated an expected and a specific nuclear staining of FOXP3 in a subset of cells which are potentially the Treg cells.

Immunohistochemistry: FoxP3 Antibody [NB100-39002] - Antibody was tested in mouse spleen using DAB with hematoxylin counterstain.

Immunohistochemistry-Paraffin: FoxP3 Antibody [NB100-39002] - FFPE bovine spleen tissue. Deparaffinization and antigen retrieval was carried out with PTlink from Dako, citrate buffer pH 6. Anti-Foxp3 antibody was used at a 1:100 dilution. Secondary reagent: Envision, from Dako. Staining: DAB and hematoxylin. IHC-P image submitted by a verified customer review.

Western Blot: FoxP3 Antibody [NB100-39002] - Jurkat Tag cells were transfected with the expression vector (vector) or with the Foxp3 expression construct (Foxp3). 24 hours after transfection, cells were lysed and analyzed by Western blot with anti-Foxp3 antiserum (1:2000 dilution).

Immunocytochemistry/Immunofluorescence: FoxP3 Antibody [NB100-39002] - Transfectants were stained with anti-Foxp3 antibody and HRPO-conjugated (upper panel) or Cy3-conjugated (lower panels) secondary antibody. Representative images (x100) are shown.

Flow Cytometry: FoxP3 Antibody [NB100-39002] - Transfectants were permeabilized and stained with anti-Foxp3 antibody and FITC-conjugated secondary antibody. Cells were analyzed by flow cytometry. Percentage of positively stained cells are shown.

Flow Cytometry: FoxP3 Antibody [NB100-39002] - An intracellular stain was performed on MCF7 cells with FoxP3 Antibody NB100-39002PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Phycoerythrin.

Dual RNAscope ISH-IHC: FoxP3 Antibody [NB100-39002] - CD8A mRNA (red) and FOXP3 protein (green) were detected in formalin-fixed paraffin-embedded tissue sections of human breast cancer. ACD's Integrated Co-Detection Workflow was performed using ACD RNAScope Probe Hs-CD8A and FoxP3 antibody at 1:400 dilution. Tissue was stained on Leica Bond RX using RNAscope (TM) 2.5 LS Reagent Kit-RED, BOND Polymer Refine Detection (DAB) and Hematoxylin, BOND Polymer Refine Red Detection and Hematoxylin and RNAscope (TM) 2.5 LS Green Accessory Pack. Tissue was counterstained with 50% hematoxylin (blue).

Dual RNAscope ISH-IHC: FoxP3 Antibody [NB100-39002] - CD4 mRNA (red) and FOXP3 protein (green) were detected in formalin-fixed paraffin-embedded tissue sections of human breast cancer. ACD's Integrated Co-Detection Workflow was performed using ACD RNAScope Probe Hs-CD4 and FOXP3 antibody at 1:200 dilution. Tissue was stained on Leica Bond RX using RNAscope (TM) 2.5 LS Reagent Kit-RED, BOND Polymer Refine Detection (DAB) and Hematoxylin, BOND Polymer Refine Red Detection and Hematoxylin and RNAscope (TM) 2.5 LS Green Accessory Pack. Tissue was counterstained with 50% hematoxylin (blue).

Effects of anaphylatoxins on transcription factor phosphorylation in ARPE-19 cells.(D) Immunofluorescence staining of FoxP3. Upper panel: Lewis rat (albino) eye with experimental uveitis; lower panel: normal rat eye. Asterisks mark immune cells infiltrating the retina from the choroid through the retinal pigment epithelium (RPE) in the eye with uveitis (clinical score of 2, histological score of 1).

FoxP3 was detected in immersion fixed MCF7 human breast cancer cell line using Rabbit anti-FoxP3 Affinity Purified Polyclonal Antibody conjugated to DyLight 488 (Catalog # NB100-39002G) (green) at 10 µg/mL overnight at 4C. Cells were stained counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunocytochemistry/ Immunofluorescence: FoxP3 Antibody - BSA Free [NB100-39002] - CCR10-expressing CD4+CD25+FOXP3+ Treg cells are enriched in the spinal cord after SCI. (A, B) Representative flow cytometry dot plots (A) & percentage (B) of CD4+CD25+FOXP3+ cells among the CD4+ T cells in the spinal cord after sham or SCI surgery. Cells were gated on 7-AAD negative population & the numbers inside plots refer to % Treg cells (n=5). (C, D) Representative histogram of CCR10 (C) & CCR3 (D) expression in CD4+CD25+FOXP3+ cells as gated in (A). (E) CD4+CD25+FOXP3+ cells in the spinal cord from sham & SCI mice were sorted out & analyzed by Western blotting to detect the expression of CCR10 & CCR3. GAPDH was used as a loading control. (F) Double immunostaining of FOXP3 (red) & CCR10 (green) in the spinal cord sections after sham or SCI surgery. The merged images are also shown. Arrows indicate cells with positive staining & colocalization. Scale bar, 100 μm. Data are mean ± SD. The statistical analysis was performed using Student’s t-test. **, P<0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31557129), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: FoxP3 Antibody - BSA Free [NB100-39002] - Effects of anaphylatoxins on transcription factor phosphorylation in ARPE-19 cells. (A–C) Protein phosphorylation of (A) CREB (Ser133); (B) FoxP3 (Ser418); & (C) FOXO1 (Ser256) was examined in ARPE-19 cells treated with C3a, C5a, or the combination of C3a/C5a at 1, 5, 10, & 15 min. Phosphorylation levels were normalized to beta -actin (relative p-protein levels), & slopes of the time course were determined. Anaphylatoxins were applied at concentrations of 300 nM (C3a) & 50 nM (C5a). Data are mean + SEM, n = 3, *p < 0.05, ***p < 0.001 (Student’s t-test). (D) Immunofluorescence staining of FoxP3. Upper panel: Lewis rat (albino) eye with experimental uveitis; lower panel: normal rat eye. Asterisks mark immune cells infiltrating the retina from the choroid through the retinal pigment epithelium (RPE) in the eye with uveitis (clinical score of 2, histological score of 1). Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fimmu.2017.00703/full), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bv

Species Glossary

Reactivity: Hu, Mu, Rt, Bv

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-39002AF350
Alexa Fluor 405	NB100-39002AF405
Alexa Fluor 488	NB100-39002AF488
Alexa Fluor 532	NB100-39002AF532
Alexa Fluor 594	NB100-39002AF594
Alexa Fluor 647	NB100-39002AF647
Alexa Fluor 700	NB100-39002AF700
Alexa Fluor 750	NB100-39002AF750
Allophycocyanin	NB100-39002APC
Biotin	NB100-39002B
CoraFluor 1	NB100-39002CL1
DyLight 350	NB100-39002UV
DyLight 405	NB100-39002V
DyLight 488	NB100-39002G
DyLight 550	NB100-39002R
DyLight 594	NB100-39002DL594
DyLight 650	NB100-39002C
DyLight 680	NB100-39002FR
DyLight 755	NB100-39002IR
FITC	NB100-39002F
HRP	NB100-39002H
Janelia Fluor 525	NB100-39002JF525
Janelia Fluor 549	NB100-39002JF549
Janelia Fluor 646	NB100-39002JF646
Janelia Fluor 669	NB100-39002JF669
PE	NB100-39002PE
PerCP	NB100-39002PCP
mFluor Violet 450 SE	NB100-39002MFV450
mFluor Violet 500 SE	NB100-39002MFV500
mFluor Violet 610 SE	NB100-39002MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-39002SS

compare

(select up to 3 products from this page to compare)

AF1759

Oct-3/4 Antibody [Unconjugated]

(76 Publications)

100 μg

AF1759

Western blot shows lysates of BG01V human embryonic stem cells and D3 mouse embryonic stem cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Oct-3/4 at approximately 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Alkaline phosphatase (ALPL) and Oct-3/4 were detected in human BG01V embryonic stem cells using 10 µg/mL Human/Mouse/Rat ALPL Monoclonal Antibody (Catalog # MAB1448) and 10 µg/mL Goat Anti-Human/Mouse Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759). Cells were incubated with primary antibodies for 3 hours at room temperature. Cells were stained for ALPL using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (pseudo-stained green; Catalog # NL007), and stained for Oct-3/4 using the NorthernLights 637-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL002). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

BG01V human embryonic stem cells were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Oct‑3/4/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Mouse Oct‑3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The nanog promoter was detected by standard PCR.

Simple Western lane view shows lysates of BG01V human embryonic stem cells and NTera‑2 human testicular embryonic carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Oct‑3/4 at approximately 50 kDa (as indicated) using 20 µg/mL of Goat Anti-Human/Mouse Oct‑3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

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

Neural induction of hPSCs. (a) Both hPSC culture and neural differentiation were performed on LN521 substrate. Cortical neurons were induced with dual SMAD inhibition, expanded in the presence of FGF2, and matured with support by a selection of neurotrophic factors. Neural progenitor cells (NPCs) could be cryopreserved at day 21 and plated at day 32 for final experiments, including microelectrode array (MEA) measurements. (b) Three different hPSC lines (one hESC line, 08/023 and two hiPSC lines, 10212.EURCCs and IMR90-4) were characterized for their efficiency in producing neuroectodermal cells in response to 12-day neural induction by dual SMAD inhibition. Cells were stained for pluripotency marker Oct4 at the pluripotent stage and after 12 days of neural induction. The presence of early neuroectodermal markers was evaluated with Sox2, FoxG1 and Pax6 staining. (c) After 21 days of differentiation, the culture contained vimentin- and Pax6-positive NPCs that could be cryopreserved. Additionally, the first Tbr2- and MAP2-positive neurons were detected at this time point. (d) The percentages of Pax6-positive cells were quantified at four time points of differentiation (mean ± s.e.m., n = 5–14, data derived from 1–3 independent differentiations). Statistical analysis was performed with the Mann-Whitney U test to compare differences between day 12 and day 46 within each hPSC line, and significant p-values are presented in the image. The scale bar is 50 µm in all images. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31748598), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, , , ,

Applications: , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	AF1759U-100UG
Alexa Fluor 405	AF1759V-100UG
Alexa Fluor 488	AF1759G-100UG
Alexa Fluor 532	AF1759X-100UG
Alexa Fluor 594	AF1759T-100UG
Alexa Fluor 647	AF1759R-100UG
Alexa Fluor 700	AF1759N-100UG
Alexa Fluor 750	AF1759S-100UG
Allophycocyanin	AF1759A
Biotin	BAF1759
CoraFluor 1	AF1759CL1
DyLight 350	AF1759D
DyLight 405	AF1759E
DyLight 488	AF1759K
DyLight 550	AF1759L
DyLight 594	AF1759M
DyLight 650	AF1759W
DyLight 680	AF1759Y
DyLight 755	AF1759Z
FITC	AF1759F
HRP	AF1759H
Janelia Fluor 525	AF1759JF525
Janelia Fluor 549	AF1759I
Janelia Fluor 646	AF1759J
Janelia Fluor 669	AF1759JF669
PE	AF1759P
PerCP	AF1759C
mFluor Violet 450 SE	AF1759MFV450
mFluor Violet 500 SE	AF1759MFV500
mFluor Violet 610 SE	AF1759MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF1759-SP

compare

(select up to 3 products from this page to compare)

NB300-320

GAPDH Antibody

(2 Reviews)

(69 Publications)

0.1 mg

NB300-320

Immunohistochemistry-Paraffin: GAPDH Antibody [NB300-320] - Staining of paraffin embedded Human Pancreas. Antibody at 2.5 ug/mL. Steamed antigen retrieval with citrate buffer pH 6, AP-staining.

Western Blot: GAPDH Antibody [NB300-320] - stai Mutants Have Reduced Levels of alpha-Tubulin, Acetylated alpha-Tubulin, and Kinesin Heavy Chain Protein. Representative western blots of total protein extracted from stai mutant third instar larvae. Heterozygous stai/+ mutant larvae exhibit mild reductions in the levels of alpha-tubulin and acetylated alpha-tubulin compared with wild type larvae. Homozygous stai mutant larvae have dramatic reductions in the levels of alpha-tubulin, acetylated alpha-tubulin and the heavy chain subunit of the microtubule motor protein kinesin compared to wildtype. Df(2L) BSC239 represents a second chromosomal deficiency that excludes the stai gene. GAPDH is used as a loading control. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0068324), licensed under a CC-BY license.

Western Blot: GAPDH Antibody [NB300-320] - Staining of HEK293 (A) and HeLa (B) cell lysate (35 ug protein in RIPA buffer). Antibody at 0.001 ug/mL. Detected by chemiluminescence. Approx 36 kDa band observed in lysates of cell line HEK293 and approx. 35 kDa in HeLa cell lysates.(calculated MW of 36.1 kDa according to Human NP_002037.2).

Immunocytochemistry/Immunofluorescence: GAPDH Antibody [NB300-320] - Immunofluorescence analysis of paraformaldehyde fixed A549 cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10 ug/mL) followed by Alexa Fluor 488 secondary antibody (2 ug/mL), showing cytoplasmic and vesicle staining. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 ug/mL) followed by Alexa Fluor 488 secondary antibody (2 ug/mL). Strong expression of the protein seen in the cytoplasm and vesicles of A549 cells.

Immunohistochemistry-Paraffin: GAPDH Antibody [NB300-320] - Upregulation of GAPDH and Tom20 in C6 gliomas. A small glioma labeled with antibodies against GAPDH. The brightness and contrast have been increased. Image collected and cropped by CiteAb from the following publication (//www.biomedcentral.com/1756-0500/8/207), licensed under a CC-BY license.

Upregulation of GAPDH & Tom20 in C6 gliomas. a A section of rat brain stained with hematoxilin & eosin to show a glioma produced by implantation of C6 cells in the right caudate nucleus. In this case, cells also grew in the region of the syringe needle track through the cortex. (b–d). A small glioma labeled by bisbenzamide (b) & with antibodies against Tom20 (c) & GAPDH (d). The brightness & contrast have been increased. e Illustration of a strip perpendicular to the rim of a glioma (labeled with bisbenzamide) & along which intensity profiles were measured. f, g Tom20 & GAPDH fluorescence along such a strip (barely visible without enhancement). The graphs show the raw intensity values given by ImageJ. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26032618), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GAPDH Antibody [NB300-320] - PAR2 causes contraction of smooth muscle cells from human prostate. (A & B) Decrease in diameter of collagen hydrogels after PAR2 activation with SLIGKV (80 µM). (C) Representative western blot & (D) densitometry showing time dependent increase in level of phosphorylated MLC20 in PSMC after PAR2 is activated. Data represent mean ± SEM of at least three independent experiments. Diameter of collagen hydrogels were measured in ImageJ (version 1.50i) & significance analyzed in Prism (version 7.04) with one‐way ANOVA followed by Tukey's multiple comparison test. *P < 0.05, **P < 0.01 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31198907), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: GAPDH Antibody [NB300-320] - Upregulation of GAPDH & Tom20 in C6 gliomas. a A section of rat brain stained with hematoxilin & eosin to show a glioma produced by implantation of C6 cells in the right caudate nucleus. In this case, cells also grew in the region of the syringe needle track through the cortex. (b–d). A small glioma labeled by bisbenzamide (b) & with antibodies against Tom20 (c) & GAPDH (d). The brightness & contrast have been increased. e Illustration of a strip perpendicular to the rim of a glioma (labeled with bisbenzamide) & along which intensity profiles were measured. f, g Tom20 & GAPDH fluorescence along such a strip (barely visible without enhancement). The graphs show the raw intensity values given by ImageJ. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26032618), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GAPDH Antibody [NB300-320] - CDC20B was the protein effector regulated by hsa-miR-633. (a) CDC20B level was up-regulated after ox-LDL treatment revealed by Western blotting. (b) CDC20B level was decreased after the treatment of hsa-miR-633 mimics revealed by Western blotting. (c) CDC20B level was increased after the treatment of hsa-miR-633 inhibitor revealed by Western blotting. (d) The luciferase reporter plasmids were constructed as illustrated. (e) Relative luciferase activities after co-transfection CDC20B WT & hsa-miR-633/Control, or CDC20B Mut & hsa-miR-633/Control were measured. (f) The expression levels of two proliferation markers, Ki67 & PCNA after the treatment of si-CDC20B were measured using Western blotting. (g) Colony formation assay was conducted to examine the proliferation abilityafter the treatment of si-CDC20B. (h) Transwell migration & invasion assays were conducted to examine the migration & invasion abilities of VSMCs after the treatment of si-CDC20B. (i) Quantification results of (g). (j) Quantification results of (h). *P < 0.05, **P < 0.01, ***P < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35212610), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Simple Western: GAPDH Antibody [NB300-320] - MiR-34a affects stroke outcomes via interacting with cytochrome c. (A) WES system image showing CYC, VDAC & GAPDH expression from hemispheres of WT & miR-34a−/− mice at 6 h post-stroke. (B) Relative CYC expression normalized to GAPDH from the data generated by the WES system. CYC level is significantly decreased in ischemic hemispheres of WT mice but no significant changes in miR-34a−/− mice between contralateral hemispheres (Control) & ischemic hemispheres (Ischemia). N = 5 per group, **p < 0.01, One-way ANOVA followed by post hoc Tuckey’s test was used for data analysis. Data are expressed as mean ± S.D. (C) Relative VDAC expression normalized to GAPDH from the data generated by the WES system. VDAC was not significantly altered in WT mice nor miR-34a−/− mice. (D) Multiplexed WES system image showing CYC & GAPDH expression from purified pCECs of WT & miR-34a−/− mice (n = 10 per group, pooled cell samples) at 6 h post-stroke. (E) Relative CYC expression by normalization to GAPDH. A 2.8 fold decrease of CYC level was observed in WT mice but no changes were observed in miR-34a−/− mice between contralateral hemispheres & ischemic hemispheres. (F) A CYC reporter was coexpressed with a miR-34a plasmid, a miR-34a mimic, a miR-34c mimic, or a plasmid control in cultured cerebral vascular endothelial cells for 24 hours. Relative firefly luciferase activity was evaluated & normalized to renilla luciferase activity. Relative firefly luciferase activity was reduced by miR-34a plasmid & miR-34a mimic. The experiment was repeated 3 times & triplicates were used for each analysis. Data represents the mean ± S.D. *p < 0.05. One-way ANOVA followed by post-hoc Tuckey’s test was used for analysis. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32094435), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GAPDH Antibody [NB300-320] - Hsa-miR-633 inhibitor could reverse the si-circ_0008896 phenotypes. (a) CCK8 assay was conducted to measure cell viability after up-regulation of hsa-miR-633. (b) The expression levels of two proliferation markers, Ki67 & PCNA after up-regulation of hsa-miR-633 were measured using Western blotting. (c) CCK8 assay was conducted to measure cell viability after the treatment of si-circ0008896, hsa-miR-633 inhibitor & si-circ0008896+ hsa-miR-633 inhibitor. (d) The expression levels of two proliferation markers, Ki67 & PCNA after the treatment of si-circ0008896, hsa-miR-633 inhibitor & si-circ0008896+ hsa-miR-633 inhibitor were measured using Western blotting. (e) Colony formation assay was conducted to examine proliferation ability number of colonies after the treatment of si-circ0008896, hsa-miR-633 inhibitor & si-circ0008896+ hsa-miR-633 inhibitor. (f) Quantification results of (e). (g) Transwell migration & invasion assays were conducted to examine the migration & invasion abilities of VSMCs after the treatment of si-circ0008896, hsa-miR-633 inhibitor & si-circ0008896+ hsa-miR-633 inhibitor. (h) Quantification results of (g). *P < 0.05, **P < 0.01, ***P < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35212610), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GAPDH Antibody [NB300-320] - Down-regulation of hsa_circ_0008896 inhibited proliferation, migration & invasion in vitro. (a) The expression level of hsa_circ_0008896 after si-circ_0008896 transfection was detected using quantitative PCR. (b) CCK8 assay was conducted to measure cell viability after down-regulation of hsa_circ_0008896. (c) The expression levels of two proliferation markers, Ki67 & PCNA after down-regulation of hsa_circ_0008896 were detected using Western blotting. (d) Colony formation assay was conducted to examine proliferation ability after down-regulation of hsa_circ_0008896. (e) The quantification results of (d). (f) Transwell migration & invasion assays were conducted to examine the migration & invasion abilities of VSMCs after down-regulation of hsa_circ_0008896. (g) The quantification results of (f). *P < 0.05, **P < 0.01, ***P < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35212610), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ca, Dr, I, Po

Species Glossary

Reactivity: Hu, Mu, Rt, Ca, Dr, I, Po

Applications:

, , , , ,

Applications: , , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Alt Catalog#

IMG-3073

Alt Catalog# IMG-3073

Conjugate:

Unconjugated

compare

(select up to 3 products from this page to compare)

NB110-40877

Perilipin-2/ADFP Antibody - BSA Free

(1 Review)

(65 Publications)

0.1 ml

NB110-40877

Formalin-fixed paraffin-embedded tissue sections of mouse intestine were probed for Perilipin-2/ADFP mRNA (ACD RNAScope Probe, catalog #577111; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog # NB110-40877) at 1:150 dilution with overnight incubation at 4 degrees Celsius followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to intestinal villi.

<b>Orthogonal Strategies Validation. </b>Simple Western lane view shows a specific band for ADFP in 0.5 mg/ml of HepG2 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

<b>Biological Strategies Validation. </b>Oleic acid increasesI2 oxidation, glucose utilization and expression of genes involved in LCFA metabolism in U138 GBM cells. Cropped western blot images and quantitation of PLIN2 and GAPDH protein levels from cell lysates. Human hepatoma (VL17A) cell line was used as a positive control (+control). Full length blots are in Supplementary Fig. A1. N=3 independent experiments in triplicate. Image collected and cropped by Citeab from the following publication (Lipid accumulation and oxidation in glioblastoma multiforme. Sci Rep (2019)) licensed under a CC-BY license.

ADFP antibody was tested in mouse liver using DAB with hematoxylin counterstain.

NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Perilipin-2/ADFP Antibody NB110-40877 at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Detection of murine ADFP in mouse liver lysate.

HepG2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-Perilipin-2/ADFP at a 1:100 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

HepG2 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-Perilipin-2/ADFP Antibody NB110-40877 at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Western Blot: Perilipin-2/ADFP Antibody - BSA Free [NB110-40877] - Western blot images (A) & quantifications (B–I) of muscle samples from WT & UCHL1 smKO mice for lipase ATGL (B), HSL (C), & MAGL (D), the fatty acid transport protein CD36 (E), the key lipid synthesis enzyme DGAT2 (F), perilipin 3 (G), perilipin 2 (H), & UCHL1 (I). n = 4-5 per group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35464088), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Perilipin-2/ADFP Antibody - BSA Free [NB110-40877] - Western blot images (A) & quantifications (B–G) of cell lysates from C2C12 myotubes with control or UCHL1 siRNA knockdown for perilipin 2 (B), perilipin 5 (C), CD36 (D), HSL (E), MAGL (F), & SDHB (G). n = 4 per group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35464088), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Perilipin-2/ADFP Antibody - BSA Free [NB110-40877] - Lack of MT4-MMP in patrolling monocytes leads to the accumulation of Mafb+AIM+ macrophages in incipient atherosclerotic plaques. a Representative images of transverse sections of aortic sinus from Ldlr–/– mice transplanted with MT4-MMP+/+ (MT4+/+) or MT4-MMP–/– (MT4–/–) BM cells & fed a HFD for 1 week; sections were labeled for Mac3 (green), Mafb (red), & AIM (white), & with Hoechst (blue; nuclei); scale bar, 10 µm. b Number of Mac3+ cells (left), Mac3+Mafb+ cells (middle), & Mac3+Mafb+AIM+ cells (right) in the plaques of BM-transplanted Ldlr–/– mice fed a HFD for 1 week. c Representative images of transverse sections of aortic sinus from BM-transplanted Ldlr–/– mice fed a HFD for 1 week; sections were labeled for Mac3 (green), Mafb (red), & adipophilin (white), & with Hoechst (blue; nuclei); scale bar, 10 µm. d Relative % of adipophilin-positive & adipophilin-negative cells within the Mac3+Mafb+ population of BM-transplanted Ldlr–/– mice (1 week on HFD); n = 7 mice per genotype in two independent experiments. Data were tested by two-tailed Student’s t-test in b & by Fisher’s exact test in d. Results are expressed as mean ± SEM. *p < 0.05 Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-03351-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Perililpin-2/ADFP was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Rabbit anti-Perililpin-2/ADFP Affinity Purified Polyclonal Antibody conjugated to Biotin (Catalog # NB110-40877B) at 5 µg/mL overnight at 4C. Cells were stained using Streptavidin conjugated to DyLight 550 (red) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Reactivity:

Hu, Mu, Rt, Bv

Species Glossary

Reactivity: Hu, Mu, Rt, Bv

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	NB110-40877AF350
Alexa Fluor 405	NB110-40877AF405
Alexa Fluor 488	NB110-40877AF488
Alexa Fluor 532	NB110-40877AF532
Alexa Fluor 594	NB110-40877AF594
Alexa Fluor 647	NB110-40877AF647
Alexa Fluor 700	NB110-40877AF700
Alexa Fluor 750	NB110-40877AF750
Allophycocyanin	NB110-40877APC
Biotin	NB110-40877B
CoraFluor 1	NB110-40877CL1
DyLight 350	NB110-40877UV
DyLight 405	NB110-40877V
DyLight 488	NB110-40877G
DyLight 550	NB110-40877R
DyLight 594	NB110-40877DL594
DyLight 650	NB110-40877C
DyLight 680	NB110-40877FR
DyLight 755	NB110-40877IR
FITC	NB110-40877F
HRP	NB110-40877H
Janelia Fluor 525	NB110-40877JF525
Janelia Fluor 549	NB110-40877JF549
Janelia Fluor 646	NB110-40877JF646
Janelia Fluor 669	NB110-40877JF669
PE	NB110-40877PE
PerCP	NB110-40877PCP
mFluor Violet 450 SE	NB110-40877MFV450
mFluor Violet 500 SE	NB110-40877MFV500
mFluor Violet 610 SE	NB110-40877MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB110-40877SS

compare

(select up to 3 products from this page to compare)

MAB1536

HIF-1 alpha Antibody (241809) [Unconjugated]

(72 Publications)

100 μg

MAB1536

Western blot shows lysates of MCF-7 human breast cancer cell line, Balb-3T3 mouse embryonic fibroblast cell line, and PC-12 rat adrenal pheochromocytoma cell line untreated (-) or treated (+) with 150 µM CoCl2for 8 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat HIF-1 alpha/HIF1A Monoclonal Antibody, followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for HIF-1 alpha/HIF1A at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

HIF-1 alpha/HIF1A was detected in immersion fixed paraffin-embedded sections of human kidney using Mouse Anti-Human/Mouse/Rat HIF-1 alpha/HIF1A Monoclonal Antibody (Catalog # MAB1536) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

<b>Genetic Strategies Validation. </b>Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HIF-1 alpha/HIF1A knockout HeLa cell line (KO) untreated (-) or treated (+) with 1 mM DFO overnight. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat HIF-1 alpha/HIF1A Monoclonal Antibody (Catalog # MAB1536) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for HIF-1 alpha/HIF1A at approximately 110 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of A549 human lung carcinoma cell line untreated (-) or treated (+) with Hypoxia (1% O2), loaded at 0.2 mg/mL. A specific band was detected for HIF-1 alpha/HIF1A at approximately 116 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse/Rat HIF-1 alpha/HIF1A Monoclonal Antibody (Catalog # MAB1536). This experiment was conducted under reducing conditions and using the12-230 kDa separation system.

<b>Genetic Strategies Validation. </b>HIF-1 alpha/HIF1A was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with DFO but is not detected in HIF-1 alpha/HIF1A knockout (KO) HeLa Human Cell Line cell line using Mouse Anti-Human/Mouse/Rat HIF-1 alpha/HIF1A alpha/HIF1A Monoclonal Antibody (Catalog # MAB1536) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Ubc13 activity is necessary for HIF1 alpha nuclear retention. (A) ChIP assays using Control IgG and HIF1 alpha antibodies were performed on cell extracts from U2OS cells pre-treated with 2μM NSC697923 for 30 min before being exposed to 1% O2 for 3 h. Immunoprecipitated DNA was analysed by qPCR using the indicated primer pairs and normalised to input genomic DNA. (B and C) U2OS cells were transfected with HA-HIF1 alpha (P402A, P564A) or empty vector and treated with 2 μM NSC697923 for 3 h. Whole cell lysates (B) or nuclear and cytoplasmic extracts (C) were prepared. Extracts were analysed by immunoblotting using the indicated antibodies. (D) Cytoplasmic and nuclear extracts were prepared from U2OS cells treated with 2 μM NSC697923 and 5 ng/ml Leptomycin B as indicated before being exposed to 1% O2 for 3 h. P-values are significant according to the Student's t-test; *P < 0.05, **P < 0.01. Image collected and cropped by CiteAb from the following publication (//academic.oup.com/nar/article/45/16/9336/3896344), licensed under a CC-BY license. Not internally tested by R&D Systems.

Increasing doses of Allopurinol reduce HIF-1 alpha levels in normoxic HFFs and HUVEC cells.A. Characterisation of HFF and HUVEC response to hypoxia. Cells were exposed to hypoxia (1% oxygen) for the indicated periods of time. At the end of incubation, protein levels were determined in whole cell extracts by immunoblot analysis using the depicted antibodies. B. Cells were treated with Allopurinol at 10, 100 and 1000 μg/ml for 17 hours. Then the cells were lysed for assessment of the indicated protein levels. Cells were treated with Etoposide (Etop) for 24 hours under normoxia. H-cells exposed to 1% O2 for 16 hours. HIF-1 alpha levels were quantified using ImageJ software and graph depicts mean and standard deviation of a minimum of three independent experiments. Anova t-test was performed and p values calculated as follows: *p<0.05; **p<0.01; ***p<0.001. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123649), licensed under a CC-BY license. Not internally tested by R&D Systems.

NQO1 enhances HIF-1 alpha expression.(a,b) RKO cells (a) and MDA-MB-231 cells (b) were exposed to 0.5% O2 for 8 h and harvested at the indicated times. Whole-cell lysates were analysed by immunoblotting for HIF-1 alpha , HIF-1 beta , NQO1 and beta -actin. SE, short exposure; LE, long exposure. (c,d) Various concentrations of pshNQO1 and pNQO1-myc-His6 (0–10 μg) were transiently transfected into RKO (c) and MDA-MB-231 (d) cells, respectively. After 48 h, the cells were exposed to 20 or 0.5% O2 for another 2 h, and then harvested. Whole-cell lysates were analysed by immunoblotting for HIF-1 alpha , NQO1 and beta -actin. EV indicates pshCont (c) and pCDNA3.1-myc-His6 (d). SE, short exposure; LE, long exposure. (e) pNQO1-myc-His6 was transiently transfected into RKO/pshNQO1 cells. After 48 h, the cells were exposed to 0.5% O2 for 2 h, and then harvested. Whole-cell lysates were analysed by immunoblotting for HIF-1 alpha , NQO1 and beta -actin. SE, short exposure; LE, long exposure. (f) Mouse embryonic fibroblast wild type and NQO1−/− cells were exposed to 20 or 0.5% O2 for 2 h and then harvested. Whole-cell lysates were analysed by immunoblotting for HIF-1 alpha , NQO1 and beta -actin. SE, short exposure; LE, long exposure. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27966538), licensed under a CC-BY license. Not internally tested by R&D Systems.

Upregulation of NQO1 correlates with poor prognosis and expression of HIF-1 alpha in colorectal cancer.(a) An Oncomine analysis of the TCGA colorectal database indicated that NQO1 expressions are elevated in colorectal cancers (n=102) compared with normal colorectal tissues (n=19). ****P<0.0001 with unpaired t-test. (b,c) An Oncomine analysis of the Staub colorectal database (b) and the Reid database (c) indicate the elevated NQO1 mRNA expression correlates with the increased colorectal cancer grade (n=11 Grade 2, n=14 Grade 3; *P<0.02 with unpaired t-test) and that elevated NQO1 mRNA expression correlates with reduced patient survival at 5 years (**P<0.01 with unpaired t-test), respectively.The gene expression spectrum indicates that colours are z-score normalized to depict relative values within rows (left panel). They cannot be used to compare values between rows. For box plots, the centre line represents the median value, box limits are at the 25th and 75th percentiles, and whiskers represent minimum and maximum values (right panel). (d) NQO1 expression correlates with poor survival in colorectal cancer data set. Analysis of the colorectal cancer data set available through Oncomine indicates a significant correlation between the high-level expression of NQO1 and poor survival in the TCGA data set (n=21 NQO1 high, n=24 NQO1 low; P=0.0001 with log-rank analysis). HR, hazard ratio; CI, confidence interval. (e) Immunohistochemical detection of HIF-1 alpha in the high-level expression of NQO1 (n=11) compared with the low-level expression of NQO1 (n=9). Positive area score of HIF-1 alpha was determined on the most characteristic areas. The positive area score of HIF-1 alpha was evaluated by determining 10 high magnification power fields ( × 40). Statistical analysis of the average score of HIF-1 alpha positive area score is shown in the right panel (P=0.0218 with unpaired t-test). Scale bar, 100 μm. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27966538), licensed under a CC-BY license. Not internally tested by R&D Systems.

Allopurinol reduces HIF-1 alpha levels independent of PHD function.A. Cells were pre-treated with Allopurinol at 10 and 1000 μg/ml for 60 minutes and then were incubated in 1% oxygen or treated with 200 μM DFX for 16 hours. Whole cell lysates were analysed by immunoblot using the indicated antibodies. B. Cells were pre-treated with Allopurinol at 1000 μg/ml for 60 minutes and then were incubated with 20μM MG132 for 3 hours. Whole cell lysates were analysed by immunoblot using the indicated antibodies. HIF-1 alpha levels were quantified using ImageJ software and graph depicts mean and standard deviation of a minimum of three independent experiments. Anova t-test was performed and p values calculated as follows: *p<0.05; **p<0.01; ***p<0.001. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123649), licensed under a CC-BY license. Not internally tested by R&D Systems.

SFN inhibits HIF-1 alpha protein accumulation.Cell lines were treated as described below, and total cell lysates subjected to immunoblot analysis. A, A549 cells were pretreated with or without SFN and MG132 (20 µM) for 1 hour. B, MCF7 cells were pretreated for 1 hour with indicated concentration of SFN, and then treated with 100µM cobalt chloride (Co+2) for an additional 5.5hours. C, 4T1 cells were pretreated for 20 min with 20 µM SFN, followed by treatment with 250µM cobalt chloride or 200µM desferroxamine as indicated for 4hours. D. 293 cells were pretreated for 20 min with indicated concentrations of SFN, followed by stimulation with 250 µM cobalt chloride for an additional 4 hours. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23166763), licensed under a CC-BY license. Not internally tested by R&D Systems.

XIAP ubiquitinates HIF1 alpha . (A) U2OS cells were transfected with siRNA oligonucleotides and His-tagged ubiquitin as indicated. Lysates were prepared and precipitated using nickel-conjugated beads followed by western blotting using the indicated antibodies. (B) HA-tagged plasmid encoding HIF1 alpha was coexpressed in HEK293 cells with His-tagged ubiquitin (Ub), FLAG-XIAP (X) or FLAG-VHL (V) as indicated. Lysates were prepared 48 h post-transfection and ubiquitinated material was recovered by incubation with nickel-conjugated beads and analysed by western blotting. (C) Biotinylated-HIF1 alpha was coexpressed in HEK293 cells with FLAG-tagged XIAP. HIF1 alpha was recovered from lysates by incubation with streptavidin-coated beads and analysed using antibodies directed against specific ubiquitin chains. (D) U2OS cells were pre-treated with 2μM NSC697923 for 30 min before being exposed to 1% O2 for 3 h. Cells were lysed in the presence of UBQ-TUBEs. Ubiquitin conjugates were purified using NiNTA agarose beads and analysed by immunoblotting using the indicated antibodies. (E) U2OS cells were pre-treated with 2 μM NSC697923 for 30 min before being exposed to 1% O2 for 3 h. Cells were lysed in the presence of TAB2-TUBEs or TAB2-AA-TUBEs as indicated. Proteins conjugated to Lys63-linked ubiquitin chains were purified using Ni-NTA agarose beads and analysed by immunoblot. Image collected and cropped by CiteAb from the following publication (//academic.oup.com/nar/article/45/16/9336/3896344), licensed under a CC-BY license. Not internally tested by R&D Systems.

SFN suppresses the HIF-1–mPGES –PGE2 axis by control of HIF-1 alpha protein expression without altering mRNA levels.A, A549 cells were pretreated with or without SFN for 30 minutes. IL1 beta (1 ng/ml) was then added and cells were cultured for another 4 hours. Total RNA was analyzed by quantitative RT-PCR. The results are expressed as fold change relative to untreated cells (mean ± SEM; n = 3). B&C, A549 cells were pretreated with or without SFN for 30 minutes. IL1 beta (1 ng/ml) was then added or not and cells were cultured for another 4 hours. Total cell lysates were subjected to immunoblot analysis (panel B) and total RNA was analyzed by quantitative RT-PCR (panel C). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23166763), licensed under a CC-BY license. Not internally tested by R&D Systems.

SFN suppresses mPGES expression and PGE2 production driven by overexpression of HIF1 alpha .A549 cells were transfected with indicated amounts of HIF-1 alpha and GFP expression vectors. The total amount of transfected DNA (4 µg) was kept constant by addition of the empty vector. The cells were then left untreated or treated with increasing concentrations of SFN for 40 hours. The total cell extracts were prepared for immunoblot analysis to evaluate protein levels. The cell culture media were used for measuring PGE2 concentration by EIA assay. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23166763), licensed under a CC-BY license. Not internally tested by R&D Systems.

Identification of histone-acetylation and HIF-destabilization in H23 cells upon co-treatment with panobinostat and cisplatin. (A) H23 cells were treated with increasing panobinostat concentrations under normoxic conditions and the amount of acetylated histone proteins H3 and H4 was detected by immunoblotting. (B) H23 cells were treated with different combinations of cisplatin or/and panobinostat both under normoxic (NOX) or hypoxic (HOX) conditions. The amounts of Ac-H3 and Ac-H4 were analyzed 24 hours later by immunoblotting. (C) After 24 hours treatment with increasing panobinostat concentrations immunoblotting for HIF-1 alpha and HIF-2 alpha was performed. (D) H23 cells were treated with cisplatin, panobinostat or with combination of both under normoxic and hypoxic conditions. After 24 hours immunoblotting for HIF-1 alpha , class II HDACs (HDAC4, 5, 6 and 7) and HDAC1 (class I) was performed. After stripping beta -actin was used as a loading control. Ac-H3 and H4, acetylated histone proteins; DMSO, vehicle control; P, panobinostat (16 nM); C, cisplatin (16 μM); P + C (16 nM panobinostat + 16 μM cisplatin); NOX, normoxia; HOX, hypoxia. (E) H23 cells were transfected with 10 nM of HIF-1 alpha siRNA and after 24 hours knock-down effects were detected by qRT-PCR and by immunoblotting. (F) H23 cells transfected with HIF-1 alpha siRNA were treated with different cisplatin concentrations for 24 hours under hypoxic conditions and cell viability was measured using the AlamarBlue® assay. Viability of untreated cells was set to 100%. Data were compared to cells transfected with control, non-silencing RNA (CTRL). ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001. (G) Protein-protein interaction between HIF-1 alpha and HDAC4 was determined by Duolink in situ proximity ligation assay and visualized by fluorescence microscopy. For negative control primary antibodies were omitted. Magnification: 200x. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25608569), licensed under a CC-BY license. Not internally tested by R&D Systems.

Increasing doses of Allopurinol reduce HIF-1 alpha levels in hypoxic HFFs, without changing HIF mRNA levels but have reduced effect in HUVEC cells.A. Cells were pre-treated with Allopurinol at 10, 100 and 1000 μg/ml for 5, 30 and 60 minutes then were incubated in 1% oxygen for 16 hours. The cells were lysed for assessment of the indicated proteins. HIF-1 alpha levels were quantified using ImageJ software and graph depicts mean and standard deviation of a minimum of three independent experiments. Anova t-test was performed and p values calculated as follows: *p<0.05; **p<0.01; ***p<0.001. B. HIF-1 alpha and HIF-2 alpha mRNA levels were analysed by quantitative PCR in HFF and HUVEC cells. Graph depicts the mean and standard deviation of a minimum of three independent experiments performed in duplicate. Anova t-test was performed and p values calculated as follows: *p<0.05; **p<0.01; ***p<0.001. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123649), licensed under a CC-BY license. Not internally tested by R&D Systems.

Generation of 3D tumor spheroid micropillar array.a Photo of 330-micropillar/well chip for effector cell-mediated cytotoxicity. The diameters of micropillar and microwell array spots are 1 and 1.9 mm, respectively. b Generation of 3D tumor spheroid by printing high-density cells (~2500 cells/250 nL) in Matrigel, followed by culturing cancer cells for up to 8 days. c Expression of HIF-1 alpha inside MiaPaCa-2 3D tumor spheroids. Confocal microscopy images of MiaPaCa-2 3D tumor spheroids stained with Hoechst 33342 and labeled with HIF-1 alpha antibody via immunofluorescence. Top views of cross-sectional images in a 3D tumor spheroid at different focal planes (top, middle, and bottom) using Imaris software. A green arrow symbol orients the viewing direction for the cross-sections reconstructed from Z stacks. d 3D view of a 3D tumor spheroid after staining the nucleus and labeling HIF1 alpha . Scale bars show 300 µm. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34290356), licensed under a CC-BY license. Not internally tested by R&D Systems.

HIF mediates GYS1 induction by hypoxia.c2c12 myoblasts were transfected with siRNA against mouse HIF1 alpha (HIF1 alpha ), an siRNA containing an irrelevant sequence (Scramble) or left untreated (none). 24 hours after transfection cells were cultured for 12 hours at 21% (Nx) or 1% oxygen (Hx) and then processed to determine mRNA (A,C), or protein (B) levels. Graphs represent normalized mRNA levels in each condition relative to the untreated normoxic cells. Data shown are the results of three independent experiments and their mean. **, p<0.01 as compared to samples transfected with irrelevant siRNA (Control) and exposed to the same oxygen tension. Numbers represent average values. (D) HIF-competent (HIF beta wt), Hepa C1, and HIF-deficient (HIF beta def), Hepa C4, cells were exposed to normoxia (Nx) or 1% oxygen (Hx) for 12 hours and processed to determine GYS1 mRNA level. The graph represents the normalized GYS1 mRNA levels in each condition relative to the normoxic Hepa C1 cells (horizontal line) from three independent experiments. **, p<0.01 as compared to HIF-competent cells exposed to the same oxygen tension. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/20300197), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , ,

Applications: , , , ,

Host:

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	FAB1536U-100UG
Alexa Fluor 405	FAB1536V-100UG
Alexa Fluor 488	FAB1536G-100UG
Alexa Fluor 532	FAB1536X-100UG
Alexa Fluor 594	FAB1536T-100UG
Alexa Fluor 647	FAB1536R-100UG
Alexa Fluor 700	FAB1536N-100UG
Alexa Fluor 750	FAB1536S-100UG
Allophycocyanin	FAB1536A
Biotin	FAB1536B
CoraFluor 1	FAB1536CL1
DyLight 350	FAB1536D
DyLight 405	FAB1536E
DyLight 488	FAB1536K
DyLight 550	FAB1536L
DyLight 594	FAB1536M
DyLight 650	FAB1536W
DyLight 680	FAB1536Y
DyLight 755	FAB1536Z
FITC	FAB1536F
HRP	FAB1536H
Janelia Fluor 525	FAB1536JF525
Janelia Fluor 549	FAB1536I
Janelia Fluor 646	FAB1536J
Janelia Fluor 669	FAB1536JF669
PE	FAB1536P
PerCP	FAB1536C
mFluor Violet 450 SE	FAB1536MFV450
mFluor Violet 500 SE	FAB1536MFV500
mFluor Violet 610 SE	FAB1536MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB1536-SP

compare

(select up to 3 products from this page to compare)

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

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

Marker

Modification

Clonality

Clone

Gene Symbol

Images Type

Isotype

Preservative

Protein Or Peptide

Research Areas

Has Sample Size

Promotional Item

Species Glossary

Primary Antibodies

NKp46/NCR1 Antibody [Unconjugated]

Caspase-1 Antibody (14F468) - BSA Free

PDX-1/IPF1 Antibody [Unconjugated]

IL-21 Antibody [Unconjugated]

PD-L1 Antibody [Unconjugated]

Noxa Antibody (114C307.1R) - BSA Free

SREBP1 Antibody (2A4) - BSA Free

FoxP3 Antibody - BSA Free

Oct-3/4 Antibody [Unconjugated]

GAPDH Antibody

Perilipin-2/ADFP Antibody - BSA Free

HIF-1 alpha Antibody (241809) [Unconjugated]