Invitrogen

HIF-2 alpha Polyclonal Antibody

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated. View Details

17 Published Figures

14 References

View all (29) HIF-2 alpha antibodies

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Datasheet

Questions & Answers

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HIF-2 alpha Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of HIF-2 alpha was performed using HepG2 cells treated with CoCl2 (300uM for 16hrs). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with HIF-2 alpha Rabbit Polyclonal Antibody (Product # PA1-16510) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the composite image showing more nuclear and less cytoplasmic localization in Hep G2 cells on treatment with CoCl2. Panel e represents untreated Hep G2 cells showing faint cytoplasmic localization. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

HIF-2 alpha Antibody in Immunocytochemistry (ICC) — Detection of HIF-2 Alpha (Green) in RCC4 cells using Product # PA1-16510. Nuclei (Blue) are counterstained with Hoechst 33258.

HIF-2 alpha Antibody in Immunohistochemistry (IHC) — Hif-2 alpha immunoreactivity in human cardiac myocytes stained with PA1-16510.

HIF-2 alpha Antibody in Western Blot (WB) — Western blot was performed using anti-HIF2A-Polyclonal Antibody (Product # PA1-16510) a 120 kDa band was observed corresponding to HIF2A was observed across the cell lines tested and increased upon Cobalt Chloride treatment in Hep G2. Whole cell Extracts of (30ug lysate) of Hep G2 (Lane 1), Hep G2 treated with Cobalt Chloride (300uM for 16hrs) (Lane 2), IMR-32 (Lane 3), tissue extracts of Rat Testis (Lane 4) and Mouse Testis (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001).The blot was probed with the primary antibody (1µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

HIF-2 alpha Antibody in Cell treatment — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot using HIF2A Polyclonal Antibody (Product # PA1-16510), shows increased expression of HIF2A in Hep G2 upon Cobalt Chloride treatment. {TM}

HIF-2 alpha Antibody — Figure 1 Arnt deletion in macrophages disrupts both HIF-1alpha and HIF -2alpha transcriptional activity. (A) PCR analysis of genomic DNA prepared from bone marrow-derived macrophages (BMDMs) and tail of LysMCre and LysMCre;Arnt fl / fl mice. The 2 lox band is the floxed allele of Arnt , and the 1 lox band indicates successful Cre-mediated recombination. (B) Western blotting of murine ARNT, HIF-1alpha, and HIF-2alpha in protein lysates prepared from LysMCre and LysMCre;Arnt fl / fl BMDMs cultured under 21% O 2 (normoxia) or 0.5% O 2 (hypoxia) for 24 h. Beta-actin was used as loading control. Densitometry was applied for quantification of HIF-1alpha and HIF-2alpha normalized to beta-actin. The numbers indicate average values of duplicated samples. (C-E) Real-time quantitative PCR (RT-qPCR) analysis of HIF target genes in LysMCre ( n = 3; mean +- s.e.m.) and LysMCre;Arnt fl / fl ( n = 3; mean +- s.e.m.) BMDMs cultured under 21% O 2 or 0.5% O 2 for 24 h. Two-way ANOVA, * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Product Details

Applications

Tested Dilution

Publications

ChIP assay (ChIP)

1:10-1:500

ELISA (ELISA)

1:100-1:2000

Flow Cytometry (Flow)

Assay-Dependent

Gel Shift (GS)

Assay-Dependent

Immunocytochemistry (ICC)

1:100

Immunofluorescence (IF)

1:100

Immunohistochemistry (Frozen) (IHC (F))

1:100

Immunohistochemistry (Paraffin) (IHC (P))

1:100

View 1 publication 1 publication

Immunoprecipitation (IP)

5 µg/1 mg lysate

Western Blot (WB)

1-2 µg/mL

View 10 publications 10 publications

Immunohistochemistry (IHC)

View 2 publications 2 publications

Miscellaneous PubMed (Misc)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Fish, Hamster, Human, Mouse, Non-human primate, Sheep, Rat

Published species

Human, Rat

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

A peptide derived from the C-terminus of mouse/human HIF-2 alpha protein.

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS

Contains

0.05% sodium azide

Storage conditions

-20°C

RRID

AB_2098236

Product Specific Information

Suggested positive control: Cos7 CoCl2-treated nuclear extract, PC12 nuclear extracts.

Target Information

HIF-2 alpha (Epas1) is a transcription factor involved in the induction of genes regulated by oxygen. HIF-2 alpha contains a basic-helix-loop-helix domain protein dimerization domain as well as a domain found in proteins in signal transduction pathways which respond to oxygen levels. It also regulates the vascular endothelial growth factor (VEGF) expression and seems to be implicated in the development of blood vessels and the tubular system of the lungs. Mutations in this gene are associated with erythrocytosis familial type 4.

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

Bioinformatics

Protein Aliases: Basic-helix-loop-helix-PAS protein MOP2; bHLHe73; Class E basic helix-loop-helix protein 73; endothelial PAS domain protein 1; Endothelial PAS domain-containing protein 1; EPAS; EPAS-1; Hif like protein; HIF-1-alpha-like factor; HIF-1alpha-like factor; HIF-2 alpha; HIF-2-alpha; HIF-2alpha; HIF-related factor; HIF1 alpha-like factor; HIF1alpha-like factor; HIF2 alpha; HIF2-alpha; HLF; HLF (HIF1alpha-like factor); HRF; hypoxia inducible factor 2, alpha subunit; hypoxia inducible factor 2a; hypoxia inducible transcription factor 2alpha; hypoxia-inducible factor 2 alpha; Hypoxia-inducible factor 2-alpha; Member of PAS protein 2; mHLF; PAS domain-containing protein 2

Gene Aliases: BHLHE73; ECYT4; EPAS; EPAS1; HIF-2alpha; HIF2A; HLF; HRF; MOP2; PASD2

UniProt ID: (Human) Q99814, (Mouse) P97481, (Rat) Q9JHS1

Entrez Gene ID: (Sheep) 443286, (Human) 2034, (Mouse) 13819, (Chinese hamster) 100771160, (Rat) 29452

Molecular Function: basic helix-loop-helix transcription factor nucleic acid binding transcription factor