Invitrogen

HIF-2 alpha Polyclonal Antibody

Advanced Verification

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

17 Published Figures

15 References

View all (30) HIF-2 alpha antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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HIF-2 alpha Antibody in Immunocytochemistry (ICC/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 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 — Cezanne and E2F1 modulate HIF2α expression. (A) HeLa and 786-O cells were transfected with control (ctrl) or Cezanne-targeting siRNAs (siCez), and whole cell lysates were prepared 48 h post-transfection, and total RNA was extracted. RT-qPCR was performed in order to analyse the mRNA levels of HIF2α and Cezanne using actin as a normalising gene (P-values are significant according to the Student's t-test; ***P<0.001). The fold-changes relative to the control are shown above the bars. (B) Schematic diagram depicting the results of the bioinformatic analysis of potential transcription factor binding sites (indicated) in the HIF2α gene promoter region. Base pairs locations are shown relative to the transcription start site (TSS). (C) 786-O cells under basal conditions and HeLa cells exposed for 24 h to 1% O2 were transfected with control or Cezanne-targeting siRNAs, and whole cell lysates were prepared 48 h post-transfection and analysed by western blotting with the antibodies indicated. (D) HeLa cells were transfected with control or E2F1-targeting siRNAs, and 48 h post-transfection, total RNA and protein extracts were prepared. RT-qPCR analysis of HIF2α, HIF1α and E2F1 mRNA was performed using actin as a normalising gene (P-values are significant according to the Student's t-test; ***P<0.001). Protein levels of HIF2α, HIF1α, HIF1β and E2F1 under the same conditions were also analysed by western blotting. The blot on the right is representative of this analysis. (E) ChIP analyses {KD}

HIF-2 alpha Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Figure 6 HIF-2alpha immunostaining in CIN model kidneys of control, CIN, and treated groups. Ratio of HIF-2alpha-positive tubular epithelial cells with immunolocalization staining in rats after modeling. According to the multiple comparison test (post-hoc test: Tukey), all possible dual comparisons were significant ( p < 0.05). QIRIAR: Quantitative immunohistochemistry score.

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1-2 µg/mL

View 10 publications 10 publications

Immunohistochemistry (IHC)

View 2 publications 2 publications

Immunohistochemistry (Paraffin) (IHC (P))

1:100

View 2 publications 2 publications

Immunohistochemistry (Frozen) (IHC (F))

1:100

Immunocytochemistry (ICC/IF)

1:100

Flow Cytometry (Flow)

Assay-Dependent

ELISA (ELISA)

1:100-1:2,000

Immunoprecipitation (IP)

5 µg/1 mg lysate

ChIP assay (ChIP)

1:10-1:500

Gel Shift (GS)

Assay-Dependent

Miscellaneous PubMed (Misc)

View 1 publication 1 publication

Product Specifications

Species Reactivity

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

Published species

Human, Mouse, 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