Invitrogen

HIF-1 beta Monoclonal Antibody (2B10)

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HIF-1 beta Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of HIF-1 beta (green) in NRK cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and blocked with 3% BSA in PBS (Product # 37525) for 30 minutes at room temperature. Cells were stained with a HIF-1 beta antibody (Product # MA1-515) at a dilution of 40 µg/mL in staining buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor 488 conjugate (Product # A28175) at a dilution of 1:1000 for 1 hour at room temperature (green). Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

HIF-1 beta Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF-7 (Lane 2), NIH/3T3 (lane 3) and PC 12 (Lane 4). The blots were probed with Anti-HIF 1 beta Mouse Monoclonal Antibody (Product # MA1-515, 1:500-1:3000 dilution) and detected by chemiluminescence Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 62-6520, 1:4000 dilution). A 84 kDa band corresponding to HIF 1 beta was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by Pierce™ Power Blotter System (22834). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

HIF-1 beta Antibody in Immunoprecipitation (IP) — Published figure using HIF-1 beta monoclonal antibody (Product # MA1-515) in Immunoprecipitation

Product Details

MA1-515

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:3,000

View 11 publications 11 publications

Immunohistochemistry (IHC)

View 2 publications 2 publications

Immunohistochemistry (Paraffin) (IHC (P))

5 µg/mL

Immunocytochemistry (ICC/IF)

Assay-dependent

View 2 publications 2 publications

Immunoprecipitation (IP)

Assay-dependent

View 2 publications 2 publications

Gel Shift (GS)

Assay-dependent

View 2 publications 2 publications

Product Specifications

Species Reactivity

Human, Mouse, Non-human primate, Rat

Published species

Chimpanzee, Fish, Human, Mouse, Non-human primate, Rat, Xenopus

Host/Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

2B10

Immunogen

Synthetic peptide corresponding to residues C N(771) S Y N N E E F P D L T M F P P F S E(789) of human ARNT.

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

purified

Storage buffer

PBS with 1mg/mL BSA

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2059441

Product Specific Information

MA1-515 detects aryl hydrocarbon (Ah) receptor nuclear translocator (ARNT) from human, mouse, rat, and primate tissues.

MA1-515 has been successfully used in Western blot, immunofluorescence, gel shift, immunocytochemistry, and immunoprecipitation procedures. By Western blot, this antibody detects an 87 kDa protein representing ARNT in COS cell extract. Immunofluorescence staining of ARNT in VT{2} cells (Hepa 1-C4T mutant deficient in ARNT function but stably transfected with the full length human ARNT cDNA) using MA1-515 yields non-nucleolar, nuclear staining only.

MA1-515 immunogen is a synthetic peptide corresponding to residues C N(771) S Y N N E E F P D L T M F P P F S E(789) of human ARNT.

Target Information

HIF-1 beta is a series of aryl hydrocarbon receptor nuclear translocator (ARNT) gene products. Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease. Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease. HIF-1 is a nuclear protein involved in mammalian oxygen homeostasis. This occurs as a posttranslational modification by prolyl hydroxylation. HIF-1 is a heterodimer composed of HIF-1 alpha and HIF-1 beta subunits. Both subunits are constantly translated. However, under normoxic conditions, human HIF-1 alpha is hydroxylated at Pro402 or Pro564 by a set of HIF prolyl hydroxylases, is polyubiquinated, and eventually degraded in proteosomes. Under hypoxic conditions, the lack of hydroxylation prevents HIF degradation and increases transcriptional activity. Therefore, the concentration of HIF-1 alpha increases in the cell. In contrast, HIF-1 beta remains stable under either condition. HIF-1 beta is a series of aryl hydrocarbon receptor nuclear translocator (ARNT) gene products. Diseases associated with HIF-1 beta dysfunction include hypoxia and renal cell carcinoma.HIF-1 beta is a series of aryl hydrocarbon receptor nuclear translocator (ARNT) gene products. Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease. Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease. HIF-1 is a nuclear protein involved in mammalian oxygen homeostasis. This occurs as a posttranslational modification by prolyl hydroxylation. HIF-1 is a heterodimer composed of HIF-1 alpha and HIF-1 beta subunits. Both subunits are constantly translated. However, under normoxic conditions, human HIF-1 alpha is hydroxylated at Pro402 or Pro564 by a set of HIF prolyl hydroxylases, is polyubiquinated, and eventually degraded in proteosomes. Under hypoxic conditions, the lack of hydroxylation prevents HIF degradation and increases transcriptional activity. Therefore, the concentration of HIF-1 alpha increases in the cell. In contrast, HIF-1 beta remains stable under either condition. HIF-1 beta is a series of aryl hydrocarbon receptor nuclear translocator (ARNT) gene products. Diseases associated with HIF-1 beta dysfunction include hypoxia and renal cell carcinoma.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Genomic Decoding of Neuronal Depolarization by Stimulus-Specific NPAS4 Heterodimers.

Cell

Brigidi GS,Hayes MGB,Delos Santos NP,Hartzell AL,Texari L,Lin PA,Bartlett A,Ecker JR,Benner C,Heinz S,Bloodgood BL

MA1-515 was used in Immunohistochemistry to show that APs and EPSPs in mouse hippocampal neurons trigger two spatially segregated and molecularly distinct induction mechanisms that lead to the expression of the ITF NPAS4.

Thu Oct 03 00:00:00 EDT 2019

A common functional regulatory variant at a type 2 diabetes locus upregulates ARAP1 expression in the pancreatic beta cell.

American journal of human genetics

Kulzer JR,Stitzel ML,Morken MA,Huyghe JR,Fuchsberger C,Kuusisto J,Laakso M,Boehnke M,Collins FS,Mohlke KL

MA1-515 was used in EMSA to study the upregulation of pancreatic beta cell ARAP1 by a functional regulatory DNA variant at a type 2 diabetes locus

Thu Feb 06 00:00:00 EST 2014

Reciprocal regulation of the basic helix-loop-helix/Per-Arnt-Sim partner proteins, Arnt and Arnt2, during neuronal differentiation.

Nucleic acids research

Hao N,Bhakti VL,Peet DJ,Whitelaw ML

MA1-515 was used in western blot to study the reciprocal regulation of the levels of Arnt and Arnt2 during neuronal development

Sat Jun 01 00:00:00 EDT 2013

Hypoxia-inducible factor and vascular endothelial growth factor are targets of dietary soy during acute stroke in female rats.

Endocrinology

Ma Y,Lovekamp-Swan T,Bekele W,Dohi A,Schreihofer DA

MA1-515 was used in western blot to study the role of HIF-1alpha and VEGF in the neuroprotective effects of soy following acute stroke

Mon Apr 01 00:00:00 EDT 2013

Effect of transient TCDD exposure on immortalized human trophoblast-derived cell lines.

Human & experimental toxicology

Fukushima K,Tsukimori K,Li D,Takao T,Morokuma S,Kato K,Seki H,Takeda S,Matsumura S,Wake N

MA1-515 was used in western blot to investigate the influence of TCDD exposure on human extravillous cytotrophoblast lines

Fri Jun 01 00:00:00 EDT 2012

Identification of residues in the N-terminal PAS domains important for dimerization of Arnt and AhR.

Nucleic acids research

Hao N,Whitelaw ML,Shearwin KE,Dodd IB,Chapman-Smith A

MA1-515 was used in western blot to investigate the role of specific amino acids for Arnt interaction with AhR

Sun May 01 00:00:00 EDT 2011

Dioxin toxicity in vivo results from an increase in the dioxin-independent transcriptional activity of the aryl hydrocarbon receptor.

PloS one

Céspedes MA,Galindo MI,Couso JP

MA1-515 was used in immunohistochemistry to study the involvemet of aryl hydrocarbon receptor in dioxin toxicity in vivo

Mon Nov 08 00:00:00 EST 2010

Amino acid substitutions in the aryl hydrocarbon receptor ligand binding domain reveal YH439 as an atypical AhR activator.

Molecular pharmacology

Whelan F,Hao N,Furness SG,Whitelaw ML,Chapman-Smith A

MA1-515 was used in western blot to investigate a novel mode of interaction between YH439 and H285Y mAhR

Tue Jun 01 00:00:00 EDT 2010

Evidence for ligand-mediated selective modulation of aryl hydrocarbon receptor activity.

Molecular pharmacology

Murray IA,Morales JL,Flaveny CA,Dinatale BC,Chiaro C,Gowdahalli K,Amin S,Perdew GH

MA1-515 was used in western blot to study the regulation of Aryl hydrocarbon receptor activity and its anti-inflammatory therapeutic potential

Mon Feb 01 00:00:00 EST 2010

The active form of human aryl hydrocarbon receptor (AHR) repressor lacks exon 8, and its Pro 185 and Ala 185 variants repress both AHR and hypoxia-inducible factor.

Molecular and cellular biology

Karchner SI,Jenny MJ,Tarrant AM,Evans BR,Kang HJ,Bae I,Sherr DH,Hahn ME

Published figure using HIF-1 beta monoclonal antibody (Product # MA1-515) in Immunoprecipitation

Wed Jul 01 00:00:00 EDT 2009

Repression of aryl hydrocarbon receptor (AHR) signaling by AHR repressor: role of DNA binding and competition for AHR nuclear translocator.

Molecular pharmacology

Evans BR,Karchner SI,Allan LL,Pollenz RS,Tanguay RL,Jenny MJ,Sherr DH,Hahn ME

Published figure using HIF-1 beta monoclonal antibody (Product # MA1-515) in Immunoprecipitation

Fri Feb 01 00:00:00 EST 2008

Repression of aryl hydrocarbon receptor (AHR) signaling by AHR repressor: role of DNA binding and competition for AHR nuclear translocator.

Molecular pharmacology

Evans BR,Karchner SI,Allan LL,Pollenz RS,Tanguay RL,Jenny MJ,Sherr DH,Hahn ME

Published figure using HIF-1 beta monoclonal antibody (Product # MA1-515) in Immunoprecipitation

Fri Feb 01 00:00:00 EST 2008

Repression of aryl hydrocarbon receptor (AHR) signaling by AHR repressor: role of DNA binding and competition for AHR nuclear translocator.

Molecular pharmacology

Evans BR,Karchner SI,Allan LL,Pollenz RS,Tanguay RL,Jenny MJ,Sherr DH,Hahn ME

Published figure using HIF-1 beta monoclonal antibody (Product # MA1-515) in Immunoprecipitation

Fri Feb 01 00:00:00 EST 2008

ARNT gene multiplicity in amphibians: characterization of ARNT2 from the frog Xenopus laevis.

Journal of experimental zoology. Part B, Molecular and developmental evolution

Rowatt AJ,DePowell JJ,Powell WH

MA1-515 was used in western blot to identify and characterize ARNT2 in Xenopus laevis

Mon Dec 15 00:00:00 EST 2003

Recruitment of dioxin receptor to active transcription sites.

Molecular biology of the cell

Elbi C,Misteli T,Hager GL

MA1-515 was used in immunocytochemistry to investigate the localization of the AhR/ARNT complex after ligand stimulation

Sat Jun 01 00:00:00 EDT 2002

Rescue of hypoxia-inducible factor-1alpha-deficient tumor growth by wild-type cells is independent of vascular endothelial growth factor.

Cancer research

Höpfl G,Wenger RH,Ziegler U,Stallmach T,Gardelle O,Achermann R,Wergin M,Kaser-Hotz B,Saunders HM,Williams KJ,Stratford IJ,Gassmann M,Desbaillets I

MA1-515 was used in western blot to investigate the role of hypoxia-inducible factor-1alpha during tumor cell proliferation.

Wed May 15 00:00:00 EDT 2002

Identification and functional characterization of hypoxia-inducible factor 2alpha from the estuarine teleost, Fundulus heteroclitus: interaction of HIF-2alpha with two ARNT2 splice variants.

The Journal of experimental zoology

Powell WH,Hahn ME

MA1-515 was used in EMSA to identify and characterize hypoxia-inducible factor 2alpha from the estuarine teleost and its interaction with ARNT2 splice variants

Mon Apr 15 00:00:00 EDT 2002

A new HIF-1 alpha variant induced by zinc ion suppresses HIF-1-mediated hypoxic responses.

Journal of cell science

Chun YS,Choi E,Yeo EJ,Lee JH,Kim MS,Park JW

MA1-515 was used in immunocytochemistry to study the effect of zinc-induced HIF-1alphaZ on HIF-1-mediated hypoxic responses.

Thu Nov 01 00:00:00 EST 2001

DNA methylation represses the expression of the human erythropoietin gene by two different mechanisms.

Blood

Yin H,Blanchard KL

MA1-515 was used in western blot to demonstrate the effect of erythropoietin CpG methylation on DNA binding, methyl-CpG binding protein recruitment and transcriptional controls.

Sat Jan 01 00:00:00 EST 2000

Up-regulation of hypoxia-inducible factor-1alpha is not sufficient for hypoxic/anoxic p53 induction.

Cancer research

Wenger RH,Camenisch G,Desbaillets I,Chilov D,Gassmann M

MA1-515 was used in western blot to investigate the mechanism for the hypoxia/anoxia up-regulation of p53

Tue Dec 15 00:00:00 EST 1998

Bioinformatics

Protein Aliases: ARNT protein; Aryl hydrocarbon receptor nuclear translocator; Aryl hydrocarbon receptor nuclear translocator 1; bHLHe2; Class E basic helix-loop-helix protein 2; Dioxin receptor, nuclear translocator; hif 1; HIF-1 beta; HIF-1-beta; HIF1 beta; HIF1-beta; hypoxia-inducible factor 1 beta; hypoxia-inducible factor 1, beta subunit; Hypoxia-inducible factor 1-beta

Gene Aliases: ARNT; Arnt1; BHLHE2; D3Ertd557e; Drnt; ESTM42; HIF-1-beta; HIF-1beta; HIF1-beta; HIF1B; HIF1BETA; mKIAA4051; TANGO; W08714

UniProt ID: (Human) P27540, (Rat) P41739, (Mouse) P53762

Entrez Gene ID: (Human) 405, (Rat) 25242, (Mouse) 11863

Molecular Function: basic helix-loop-helix transcription factor