Recombinant

Recombinant Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)

Overview

  • Product name

    Anti-HIF-2-alpha antibody [BL-95-1A2]
    See all HIF-2-alpha primary antibodies
  • Description

    Rabbit monoclonal [BL-95-1A2] to HIF-2-alpha
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IP, CHIPseq, ICC, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human HIF-2-alpha aa 400-450. The exact sequence is proprietary. NP_001421.2 and Gene ID 2034.
    Database link: Q99814

  • General notes

    This product is sold under License from Bethyl Laboratories, Inc.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab243861 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100 - 1/500.

Permeabilization with Triton-X 100 is recommended for formaldehyde-fixed cells.

IP Use at an assay dependent concentration.

Use 5-20µl/mg lysate.

CHIPseq Use a concentration of 10-40 - 1 µl/chromatin.
ICC 1/200 - 1/1000.

Permeabilization with Triton-X 100 is recommended for formaldehyde-fixed cells.

IHC-P 1/200 - 1/1000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Antigen retrieval with citrate buffer pH 6.0 for 20 minutes using a pressure cooker and overnight incubations are recommended.

WB 1/1000.

Target

  • Function

    Transcription factor involved in the induction of oxygen regulated genes. Binds to core DNA sequence 5'-[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters. Regulates the vascular endothelial growth factor (VEGF) expression and seems to be implicated in the development of blood vessels and the tubular system of lung. May also play a role in the formation of the endothelium that gives rise to the blood brain barrier. Potent activator of the Tie-2 tyrosine kinase expression. Activation seems to require recruitment of transcriptional coactivators such as CREBPB and probably EP300. Interaction with redox regulatory protein APEX seems to activate CTAD.
  • Tissue specificity

    Expressed in most tissues, with highest levels in placenta, lung and heart. Selectively expressed in endothelial cells.
  • Involvement in disease

    Defects in EPAS1 are the cause of erythrocytosis familial type 4 (ECYT4) [MIM:611783]. ECYT4 is an autosomal dominant disorder characterized by increased serum red blood cell mass, elevated hemoglobin concentration and hematocrit, and normal platelet and leukocyte counts.
  • Sequence similarities

    Contains 1 basic helix-loop-helix (bHLH) domain.
    Contains 1 PAC (PAS-associated C-terminal) domain.
    Contains 2 PAS (PER-ARNT-SIM) domains.
  • Post-translational
    modifications

    In normoxia, is probably hydroxylated on Pro-405 and Pro-531 by EGLN1/PHD1, EGLN2/PHD2 and/or EGLN3/PHD3. The hydroxylated prolines promote interaction with VHL, initiating rapid ubiquitination and subsequent proteasomal degradation. Under hypoxia, proline hydroxylation is impaired and ubiquitination is attenuated, resulting in stabilization.
    In normoxia, is hydroxylated on Asn-847 by HIF1AN thus probably abrogating interaction with CREBBP and EP300 and preventing transcriptional activation.
    Phosphorylated on multiple sites in the CTAD.
    The iron and 2-oxoglutarate dependent 3-hydroxylation of asparagine is (S) stereospecific within HIF CTAD domains.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Basic helix loop helix PAS protein MOP2 antibody
    • Basic-helix-loop-helix-PAS protein MOP2 antibody
    • bHLHe73 antibody
    • Class E basic helix-loop-helix protein 73 antibody
    • ECYT4 antibody
    • Endothelial PAS domain containing protein 1 antibody
    • Endothelial pas domain protein 1 antibody
    • Endothelial PAS domain-containing protein 1 antibody
    • EPAS 1 antibody
    • EPAS-1 antibody
    • EPAS1 antibody
    • EPAS1_HUMAN antibody
    • HIF 1 alpha like factor antibody
    • HIF 2 alpha antibody
    • HIF-1-alpha-like factor antibody
    • HIF-2-alpha antibody
    • HIF2-alpha antibody
    • HIF2A antibody
    • HLF antibody
    • Hypoxia inducible factor 2 alpha antibody
    • Hypoxia inducible factor 2 alpha subunit antibody
    • Hypoxia-inducible factor 2-alpha antibody
    • Member of PAS protein 2 antibody
    • Member of pas superfamily 2 antibody
    • MOP 2 antibody
    • MOP2 antibody
    • PAS domain-containing protein 2 antibody
    • PASD2 antibody
    see all

Images

  • Localization of human HIF2-alpha binding sites in immunoprecipitates from subcutaneous tumor 786-O cells by ChIP-Seq using ab243861.
  • Formaldehyde-fixed HepG2 (human liver hepatocellular carcinoma cell line) cells untreated (Left) and treated with CoCl2 (right), labeling HIF-2 alpha (Red) using ab243861 at 1/100 dilution. DyLight® 594-conjugated goat anti-rabbit IgG was used as the secondary antibody at 1/100 dilution. Phalloidin Alexa Fluor® 488conjugated (green) was used as the counterstain.
  • Formalin-fixed, paraffin-embedded HepG2 (human liver hepatocellular carcinoma cell line) cells labeling HIF-2 alpha using ab243861 at 1/400 dilution in ICC/IF analysis. A HRP-conjugated goat-anti rabiit IgG was used as the secondary. DAB staining.
  • Formalin-fixed, paraffin-embedded human renal cell carcinoma tissue stained for HIF-2 alpha using ab243861 at 1/400 dilution in immunohistochemical analysis. A HRP-conjugated goat anti-rabbit IgG was used as the secondary. DAB staining.
  • HIF-2 alpha was immunoprecipitated from 1 mg HepG2 whole cell lysate with ab243861 at 20 µL per reaction. Western blot was performed on the immunoprecipitate using ab243861 at 1/1000 dilution.

    Lane 1: ab243861 IP in HepG2 cell lysate treated with 200 µM CoCl2.

    Lane 2: ab243861 IP in HepG2 whole cell lysate Mock treated.

    Lane 3: Contol IgG in HepG2 cell lysate treated with 200 µM CoCl2.

  • Detection of human HIF2-alpha by WB of Hep-G2, HEK293T, and 786-O lysate treated with 200 M CoCl2 (+) or mock treated (-) using ab243861. Secondary: HRP-conjugated goat anti-rabbit IgG.

References

ab243861 has not yet been referenced specifically in any publications.

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