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  1. Link

    hif-2-alpha-antibody-ab199.pdf

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Cardiovascular Hypoxia HIF
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Anti-HIF-2-alpha 抗体 (ab199)

  • Datasheet
  • SDS
Reviews (11)Q&A (15)References (117)

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Western blot - Anti-HIF-2-alpha antibody (ab199)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-2-alpha antibody (ab199)
  • Western blot - Anti-HIF-2-alpha antibody (ab199)

Key features and details

  • Rabbit polyclonal to HIF-2-alpha
  • Suitable for: IHC-P, WB
  • Reacts with: Human, African green monkey
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-HIF-2-alpha antibody
    HIF-2-alpha 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to HIF-2-alpha
  • 由来種

    Rabbit
  • 特異性

    Specific for HIF-2-alpha / EPAS 1. Does not cross react with HIF-1-alpha.
  • アプリケーション

    適用あり: IHC-P, WBmore details
  • 種交差性

    交差種: Human, African green monkey
  • 免疫原

    Synthetic peptide corresponding to Mouse HIF-2-alpha aa 632-646.
    Sequence:

    GRSNTQWPPDPPLHF

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • ポジティブ・コントロール

    • WB: Hypoxic HeLa lysate; COS7 nuclear extracts IHC: Human cardiac myocytes
  • 特記事項

    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

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • バッファー

    pH: 7.40
    Preservative: 0.05% Sodium azide
    Constituent: 99% PBS
  • Concentration information loading...
  • 精製度

    Affinity purified
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • 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

関連製品

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

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

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab199の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
IHC-P (1)
1/100.
WB (9)
1/200 - 1/1000. Detects a band of approximately 118 kDa (predicted molecular weight: 100 kDa). The following should be taken into consideration: sufficient hypoxia induction is necessary; use nuclear extracts for cleaner results; use positive/negative controls to see which band is up-regulated; HIF2a degrades rapidly (fast sample preparation, protease inhibitors!).
特記事項
IHC-P
1/100.
WB
1/200 - 1/1000. Detects a band of approximately 118 kDa (predicted molecular weight: 100 kDa). The following should be taken into consideration: sufficient hypoxia induction is necessary; use nuclear extracts for cleaner results; use positive/negative controls to see which band is up-regulated; HIF2a degrades rapidly (fast sample preparation, protease inhibitors!).

ターゲット情報

  • 機能

    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.
  • 組織特異性

    Expressed in most tissues, with highest levels in placenta, lung and heart. Selectively expressed in endothelial cells.
  • 関連疾患

    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.
  • 配列類似性

    Contains 1 basic helix-loop-helix (bHLH) domain.
    Contains 1 PAC (PAS-associated C-terminal) domain.
    Contains 2 PAS (PER-ARNT-SIM) domains.
  • 翻訳後修飾

    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.
  • 細胞内局在

    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
  • 参照データベース

    • Entrez Gene: 2034 Human
    • Omim: 603349 Human
    • SwissProt: Q99814 Human
    • Unigene: 468410 Human
    • 別名

      • 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

    画像

    • Western blot - Anti-HIF-2-alpha antibody (ab199)
      Western blot - Anti-HIF-2-alpha antibody (ab199)
      Anti-HIF-2-alpha antibody (ab199) + Hypoxic HeLa cell lysate at 0.5 mg/ml

      Performed under reducing conditions.

      Predicted band size: 100 kDa



      Detection of HIF-2-alpha in hypoxic HeLa lysate using ab199, reducing conditions using the 12-230 kDa separation system.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-2-alpha antibody (ab199)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-2-alpha antibody (ab199)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of huaman cardiac myocytes labeling HIF-2-alpha with ab199 at 1/100 dilution.

    • Western blot - Anti-HIF-2-alpha antibody (ab199)
      Western blot - Anti-HIF-2-alpha antibody (ab199)
      All lanes : Anti-HIF-2-alpha antibody (ab199)

      Lane 1 : Cobalt chloride treated COS7 nuclear extracts
      Lane 2 : Untreated COS7 nuclear extracts

      Predicted band size: 100 kDa

    プロトコール

    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    データシートおよび資料

    • SDS download

    • Datasheet download

      Download

    参考文献 (117)

    ab199 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab199 は 117 報の論文で使用されています。

    • Macias D  et al. Targeting HIF2a-ARNT hetero-dimerisation as a novel therapeutic strategy for pulmonary arterial hypertension. Eur Respir J 57:N/A (2021). PubMed: 32972983
    • Pantazopoulou V  et al. Hypoxia-Induced Reactivity of Tumor-Associated Astrocytes Affects Glioma Cell Properties. Cells 10:N/A (2021). PubMed: 33802060
    • Niklasson CU  et al. Hypoxia inducible factor-2a importance for migration, proliferation, and self-renewal of trunk neural crest cells. Dev Dyn 250:191-236 (2021). PubMed: 32940375
    • Chan CD  et al. Co-localisation of intra-nuclear membrane type-1 matrix metalloproteinase and hypoxia inducible factor-2a in osteosarcoma and prostate carcinoma cells. Oncol Lett 21:158 (2021). PubMed: 33552276
    • Lyu Q  et al. Intermittent short-duration reoxygenation protects against simulated high altitude-induced pulmonary hypertension in rats. FASEB J 35:e21212 (2021). PubMed: 33230951
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 26 Abreviews or Q&A

    Western blot abreview for Anti-HIF-2-alpha antibody - ChIP Grade

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (10% SDS PAGE)
    Sample
    Mouse Tissue lysate - whole (Mouse brain)
    Specification
    Mouse brain
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jun 02 2014

    Western blot abreview for Anti-HIF2 alpha antibody - ChIP Grade

    Inconclusive
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Heart, Left Ventricle)
    Loading amount
    40 µg
    Specification
    Heart, Left Ventricle
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Dawn Bowles

    Verified customer

    投稿 Jan 10 2013

    Western blot abreview for Anti-HIF-2-alpha antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Leukemia T cell line)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    30 µg
    Treatment
    5uM MG132 for 6 hours
    Specification
    Leukemia T cell line
    Blocking step
    (agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 14 2021

    Western blot abreview for Anti-HIF-2-alpha antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Leukemia T cell line)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    31 µg
    Treatment
    5uM MG132 for 6 hours
    Specification
    Leukemia T cell line
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 14 2021

    Western blot abreview for Anti-HIF-2-alpha antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (cell line)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    70 µg
    Treatment
    1% oxygen
    Specification
    cell line
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 14 2021

    Western blot abreview for Anti-HIF-2-alpha antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (cell line)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    70 µg
    Treatment
    1% oxygen
    Specification
    cell line
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 14 2021

    Western blot abreview for Anti-HIF-2-alpha antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293T)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    HEK293T
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 10 2021

    ChIP abreview for Anti-HIF-2-alpha antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (Kidney (Clear Cell Renal Carcinoma))
    Negative control
    Parental clear cell renal carcinoma cells (786-O).
    Specification
    Kidney (Clear Cell Renal Carcinoma)
    Detection step
    Other
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 1% Formaldehyde
    Positive control
    Metastatic clear cell renal carcinoma cells (M1A).
    Read More

    Paulo Rodrigues

    Verified customer

    投稿 Aug 17 2018

    Question


    the customer would like to ask some more clarification:
    - Which is the the active form of HIF-2a (the band at 96 KDa or the other band at 120 kDa)?
    - Which are the post-translational modifications (phosphorilations acetylation, methylation, glycosylation, sumoylation, ubiquitination, etc.)?

    Thanks in advance and kind regards

    Read More

    Abcam community

    Verified customer

    Asked on Nov 20 2012

    Answer

    Thank you for your reply.

    As discussed, the HIF1 and 2 alpha subunits have been shown to be present in forms between 96 and 120 kDa through postranslational modifications. This has been studied in more depth with HIF1-alpha than HIF2-alpha. For example, in the following study they found a band at ˜90 kDa and one at ˜116 kDa in renal cell carcinoma cell line:

    http://clincancerres.aacrjournals.org/content/11/3/1129.full

    They found the ratio between the two bands differed depending on the cell lines tested.

    This topic can be reviewed in more detail from literature but I believe both forms are active.

    I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

    Read More

    Abcam Scientific Support

    Answered on Nov 20 2012

    Question

    Dear Sirs,
    a customer of ours has bought your ab199 and found by WB two bands (one at 95kDa and one at 120kDa).
    In your datasheet it is indicated: "For instance, unprocessed HIF2 is 96 KDa while the full post-translationally modified form is 118 KDa or larger".
    The customer would like to know what do you mean with "unprocessed HIF2" and moreover which are the full post-translationally modified forms?

    Thanks in advance and kind regards

    Read More

    Abcam community

    Verified customer

    Asked on Nov 16 2012

    Answer

    Many thanks for contacting us and your interest in our products.

    Both the HIF-1alpha and HIF-2alpha have been to show this characteristics. Whilst the expressed protein would be expected at ˜96 kDa, this can be post-translationally modified, increasing the protein size. Bands are therefore commonly observed between 96 kDa to 120 kDa.

    The level of post-translational modifications can be as a result of the compounds used to increase HIF production as well as the cell strains probed.

    I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

    Read More

    Abcam Scientific Support

    Answered on Nov 16 2012

    1-10 of 26 Abreviews or Q&A

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