Anti-HIF-2-alpha 抗体 (ab199)

Reviews (7)Q&A (15)References (99)
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
  • 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

  • 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

  • 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

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 (5)
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.
  • Information by UniProt

  • 参照データベース

    • 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

    参考文献 (99)

    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 99 報の論文で使用されています。

    • Du X  et al. Hypoxia-Inducible Factor 1a and 2a Have Beneficial Effects in Remote Ischemic Preconditioning Against Stroke by Modulating Inflammatory Responses in Aged Rats. Front Aging Neurosci 12:54 (2020). PubMed: 32210788
    • Grassi ES  et al. Hypoxia-induced release, nuclear translocation, and signaling activity of a DLK1 intracellular fragment in glioma. Oncogene 39:4028-4044 (2020). PubMed: 32205867
    • Yang T  et al. Low-Intensity Pulsed Ultrasound Alleviates Hypoxia-Induced Chondrocyte Damage in Temporomandibular Disorders by Modulating the Hypoxia-Inducible Factor Pathway. Front Pharmacol 11:689 (2020). PubMed: 32477144
    • Jia W  et al. Gadofullerene nanoparticles for robust treatment of aplastic anemia induced by chemotherapy drugs. Theranostics 10:6886-6897 (2020). PubMed: 32550910
    • Nusblat LM  et al. Gene silencing of HIF-2a disrupts glioblastoma stem cell phenotype. Cancer Drug Resist 3:199-208 (2020). PubMed: 32566921
    View all Publications for this product

    レビューと Q&A

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    1-10 of 22 Abreviews or Q&A

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

     Average
    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
    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
    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
    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
    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

    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
    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

    Question

    Thank you for your fast reply. The old PO number was: If you are going to send a replacement product, please send it to this free-of-charge PO number:

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement through Cedarlane of ab109616.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Question

    Amount: 200ug (Mouse lung tissue, animals in chronic hypoxia for 3 weeks) Blocking conditions: 5% milk in 1x TBST, 1hr, Detection method: secondary donkey anti-rabbit antibody Electrophoresis: 8% gel Transfer: Wet/2hrs. Primary: 1:500, overnight (~20hrs), 4oC. Secondary: 1 hr, room temp, 1:2500 Wash steps: three TBST after primary, three after secondary, one PBS I did not detect any bands at the correct size at all (some band at 60kDa). I did this western at the same time as for HIF1a (product: ab2185) I got ALOT of background (I am optimizing now for HIF1a). However, I dont see how this antibody will work since I got 200ug of protein loaded and 1:500 primary and very high secondary (1:2500)!! I also did the beta-actin staining, and it works fine on the same membrane (without stripping) In your sheet, it mentions that the lung is most abundant in the HIF-2a protein!! If possible, instead of a refund, I would like to get this antibody instead: ab109616. Thank you

    Read More
    Answer

    Thank you for contacting us.

    I am very sorry to hear that this product did not work as specified on our datasheet and would like to thank you for the information which you have provided. This product is covered by our Abpromise guarantee, provided that it has been purchased within the last six months. In order to send to you a replacement product I will need your Abcam confirmation number or the PO used to purchase this product from Abcam.

    I hope this information is helpful to you and look forward to your response. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

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    Question

    Lot number: GR37292-1

    Inquiry: I had ordered anti-HIF-2 alpha antibody with product code#ab199. The order number is *****.
    Unfortunately, the performance was not satisfactory. There are two problems. Firstly, we are not always detecting a band of the correct size and secondly, we always obtain a lot of background noise.
    Below, there is a description of the anti-HIF-2 alpha Western Blot procedure that we used, in order to find out if the problem is due to any of the protocol’s stages.
    We prepared whole cell lysates. The cell lines used were Huh7 and HepG2 and U87. Cells were treated under 1% hypoxia for 8 and 16 hours.
    1. We run the proteins (50μg and 100μg) on SDS 8% polyacrylamide gel.
    2. Transfered to nitrocellulose membranes. The transfer was verified by ponceau red staining.
    3. Blocked membranes for 1.5 hours with 1x western wash buffer (PBS1x + TWEEN) containing 3% BSA. (We have used the same blocking conditions with other antibodies such as anti-HIF1α, anti- actin, anti- tubulin, anti-P- MAPK, etc, so we are pretty sure that it is working).
    4. Washed with 1x western wash (PBS1x + TWEEN) for 30 minutes at RT (3 x 10 minutes).
    5. Incubated membranes with the antibody HIF2α (#ab199) 1:500 in 1x western wash 1 % BSA overnight at 4Co.
    6. Washed with 1x western wash (PBS1x + TWEEN) for 30 minutes at RT (3 x 10 minutes).
    7. Incubated membranes with the second antibody (anti-rabbit 1:1000 from Cell Signaling) in 1x western wash (PBS1x + TWEEN) 5% non fat milk.
    8. Washed with 1x western wash (PBS1x + TWEEN) for 30 minutes at RT (3 x 10 minutes).
    9. For the exposure we used chemiluminescense. We also attach some western blot films.
    Also, I would like to mention that we use the ab199 antibody (different Lot numbers) since 2008 and we haven’t any problem until now. Would you be willing to send us a small amount from a different Lot? Thanks in advance for your time.

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    Answer

    Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.
    Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty lot.
    As requested, I have issued a free of charge replacement with a new lot of ab199, the order number is **********.
    To check the status of the order please contact our Customer Service team and reference this number.
    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
    I wish you the best of luck with your research.

    Read More

    Question

    The ab199 replacement vial that I received recently turned also out not to work. I have attached a word document with all the details.

    During the last years, I have bought two vials of this antibody that have been working beautifully and since a lot of my data is based on this antibody, I’m really keen to continue with the same antibody. Do you have any lot in stock that you know is behaving well (for western blot) that I could get this non-working vial replaced with?

    I would also like to inform you, that If it is possible to deliver an ab199 of the same quality as I’ve experienced before, my plan is to order in two several vials of the same lot and store in -20 (I know it is not recommended to freeze the antibody but I’m willing to take the risk) for future usage.

    Read More
    Answer

    Thank you for confirming these details and for your cooperation.

    The details provided enable us to closely monitor the quality of our products.

    I am sorry the replacementdid not perform as stated on the datasheet.

    As requested, I have issued a free of charge replacement with a new lot. The order number is 1100174.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    1-10 of 22 Abreviews or Q&A

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