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Recombinant

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

Submit a review Submit a question References (5)
ChIP-sequencing - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
  • Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
  • Immunocytochemistry - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
  • Immunoprecipitation - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
  • Western blot - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
  • Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [BL-95-1A2] to HIF-2-alpha
  • Suitable for: ICC/IF, IP, ChIP-sequencing, ICC, IHC-P, WB
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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, ChIP-sequencing, 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

The Abpromise guarantee

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.
ChIP-sequencing
Use a concentration of 10-40 - 30 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.
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.
ChIP-sequencing
Use a concentration of 10-40 - 30 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

    • ChIP-sequencing - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      ChIP-sequencing - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)

      Chromatin from subcutaneous human tumor 786-O cells was immunoprecipitated with anti-HIF-2-alpha antibody ab243861 and analyzed by
      DNA sequencing. The figure illustrates the peak distribution of HIF-2-alpha binding within a 250 Kb region of chromosome 11 as detected
      using anti-HIF-2-alpha antibody ab243861. ChIP-seq validation performed by Active Motif, Carlsbad, CA.

    • Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [BL-95-1A2] (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.
    • Immunocytochemistry - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      Immunocytochemistry - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      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.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      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.
    • Immunoprecipitation - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      Immunoprecipitation - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      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.

    • Western blot - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      Western blot - Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      All lanes : Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861) at 1/1000 dilution

      Lane 1 : HepG2 whole cell lysate treated with 200uM CoCl2
      Lane 2 : HepG2 whole cell lysate mock treated
      Lane 3 : HEK293T whole cell lysate treated with 200uM CoCl2
      Lane 4 : HEK293T whole cell lysate mock treated
      Lane 5 : 786-O whole cell lysate mock treated

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG
    • Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)
      Anti-HIF-2-alpha antibody [BL-95-1A2] (ab243861)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (5)

    Publishing research using ab243861? Please let us know so that we can cite the reference in this datasheet.

    ab243861 has been referenced in 5 publications.

    • Lin JX  et al. m6A methylation mediates LHPP acetylation as a tumour aerobic glycolysis suppressor to improve the prognosis of gastric cancer. Cell Death Dis 13:463 (2022). PubMed: 35568711
    • Wang Z  et al. Hypoxia-induced PLOD1 overexpression contributes to the malignant phenotype of glioblastoma via NF-?B signaling. Oncogene N/A:N/A (2021). PubMed: 33420370
    • Yin R & Liu S SHARPIN regulates the development of clear cell renal cell carcinoma by promoting von Hippel-Lindau protein ubiquitination and degradation. Cancer Sci 112:4100-4111 (2021). PubMed: 34339558
    • Liu X  et al. HIF-1-regulated expression of calreticulin promotes breast tumorigenesis and progression through Wnt/β-catenin pathway activation. Proc Natl Acad Sci U S A 118:N/A (2021). PubMed: 34706936
    • Duan X & Qin G Notch inhibitor mitigates renal ischemia-reperfusion injury in diabetic rats. Mol Med Rep 21:583-588 (2020). PubMed: 31974606

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