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    hif-2-alpha-antibody-ep190b-ab8365.pdf

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Cardiovascular Hypoxia HIF
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Anti-HIF-2-alpha antibody [ep190b] (ab8365)

  • Datasheet
  • SDS
Reviews (5)Q&A (18)References (66)

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Western blot - Anti-HIF-2-alpha antibody [ep190b] (ab8365)
  • Western blot - Anti-HIF-2-alpha antibody [ep190b] (ab8365)

Key features and details

  • Mouse monoclonal [ep190b] to HIF-2-alpha
  • Suitable for: WB
  • Reacts with: Human
  • Isotype: IgG1

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Primary
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Anti-HIF-1 alpha antibody [H1alpha67] (ab1)
Secondary
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Goat Anti-Mouse IgG H&L (HRP) (ab205719)

View more associated products

Overview

  • Product name

    Anti-HIF-2-alpha antibody [ep190b]
    See all HIF-2-alpha primary antibodies
  • Description

    Mouse monoclonal [ep190b] to HIF-2-alpha
  • Host species

    Mouse
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Rat
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Hypoxic A549 and HeLa cell lysate.
  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    ep190b
  • Myeloma

    NS1
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

    • Cardiovascular
    • Hypoxia
    • HIF
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • HLH
    • Epigenetics and Nuclear Signaling
    • Cardiovascular/Immune
    • Hypoxia
    • HIF
    • Cancer
    • Invasion/microenvironment
    • Hypoxia
    • HIF
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Nucleotide metabolism
    • Molecular processes
    • Mitochondrial transcription
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab8365 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
WB (3)
Use a concentration of 1 µg/ml. Predicted molecular weight: 100 kDa.

Abcam recommends using a 3% milk block with this product.

Notes
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 100 kDa.

Abcam recommends using a 3% milk block with this product.

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.
  • Target information above from: UniProt accession Q99814 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2034 Human
    • Entrez Gene: 29452 Rat
    • Omim: 603349 Human
    • SwissProt: Q99814 Human
    • SwissProt: Q9JHS1 Rat
    • Unigene: 468410 Human
    • Unigene: 55138 Rat
    • 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

    • Western blot - Anti-HIF-2-alpha antibody [ep190b] (ab8365)
      Western blot - Anti-HIF-2-alpha antibody [ep190b] (ab8365)
      All lanes : Anti-HIF-2-alpha antibody [ep190b] (ab8365) at 1/500 dilution

      Lane 1 : HeLa whole cell lysate
      Lane 2 : HeLa treated with 0.5mM DFO whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP conjugated Goat Anti-Mouse IgG (H+L) at 1/10000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 100 kDa
      Observed band size: 100 kDa


      Exposure time: 20 minutes


      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab8365 overnight at 4°C. Antibody binding was detected using a Goat anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ab133406.

    • Western blot - Anti-HIF-2-alpha antibody [ep190b] (ab8365)
      Western blot - Anti-HIF-2-alpha antibody [ep190b] (ab8365)

      HIF-2-alpha detected in hypoxic Human lysate using ab8365. Lane 1: normoxic A549 lysate control, lane 2: hypoxic A549 lysate.

    Protocols

    • Western blot protocol for HIF-2 alpha
    • Immunohistochemistry for HIF2 alpha

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (66)

    Publishing research using ab8365? Please let us know so that we can cite the reference in this datasheet.

    ab8365 has been referenced in 66 publications.

    • Morin A  et al. TET-Mediated Hypermethylation Primes SDH-Deficient Cells for HIF2a-Driven Mesenchymal Transition. Cell Rep 30:4551-4566.e7 (2020). PubMed: 32234487
    • Rodrigues T  et al. GLP-1 improves adipose tissue glyoxalase activity and capillarization improving insulin sensitivity in type 2 diabetes. Pharmacol Res 161:105198 (2020). PubMed: 32942016
    • Guo X  et al. Whole-exome sequencing and immunohistochemistry findings in von Hippel-Lindau disease. Mol Genet Genomic Med 7:e880 (2019). PubMed: 31317677
    • Cho C  et al. Cirsium japonicum var. maackii and apigenin block Hif-2a-induced osteoarthritic cartilage destruction. J Cell Mol Med 23:5369-5379 (2019). PubMed: 31148341
    • Polívka J  et al. IDH1 mutation is associated with lower expression of VEGF but not microvessel formation in glioblastoma multiforme. Oncotarget 9:16462-16476 (2018). IHC-P ; Human . PubMed: 29662659
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-HIF-2-alpha antibody [ep190b]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Kidney)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    Kidney
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 12 2019

    Western blot abreview for Anti-HIF-2-alpha antibody [ep190b]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing (4-12% acrylamide)
    Sample
    Human Cell lysate - whole cell (Renal cancer cell line)
    Specification
    Renal cancer cell line
    Blocking step
    Li-cor Blocking Buffer PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 28 2014

    Western blot abreview for Anti-HIF2 alpha antibody [ep190b]

    Good
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK 293)
    Loading amount
    20 µg
    Specification
    HEK 293
    Gel Running Conditions
    Non-reduced Denaturing (7.5)
    Blocking step
    Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 23 2011

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