製品の概要

  • 製品名Anti-HIF-2-alpha antibody - ChIP Grade
    HIF-2-alpha 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to HIF-2-alpha - ChIP Grade
  • 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: Flow Cyt, IHC-Fr, ChIP, IP, ICC/IF, IHC-P, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

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

    GRSNTQWPPDPPLHF

  • ポジティブ・コントロール
    • CoCl2-treated Cos7 nuclear extract or hypoxic/normoxic PC12 nuclear extracts. Under normoxic conditions HIF-2 alpha has a short half-life. It is largely undetectable in cells or tissues grown under normoxic conditions. It is stabilized only at O2 concentrations below 5% and upon stabilization under hypoxic conditions HIF-2 translocate to the nucleus. Therefore we recommend western blots using nuclear extracts and running Hypoxia treated samples as positive control (ab180880).

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab199 in the following tested applications.

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

アプリケーション Abreviews 特記事項
Flow Cyt Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration. PubMed: 19897487
IP Use at 5-1 µg/mg of lysate.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/100.
EMSA Use at an assay dependent concentration.
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

Anti-HIF-2-alpha antibody - ChIP Grade 画像



  • Predicted band size : 100 kDa

    Analysis on normoxic and hypoxic nuclear rat cell lysates.

  • ab199 staining human adrenal tissue sections by IHC-P.  Sections were PFA fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking in 10% serum for 10 minutes at 25°C.  The primary anitbody was diluted 1/500 and incubated with the sample for 1 hour at 25°C.  A biotinylated goat anti-rabbit antibody was used as the secondary.

    See Abreview



  • Performed under reducing conditions.

    Predicted band size : 100 kDa

    Image from PLoS One. 2012; 7(1): e29876. Fig3A, doi: 10.1371/journal.pone.0029876

    HIF-2α levels in HIF-2α−/− and WT mouse kidneys.

    HIF-2α−/− mice or their wild-type littermates were exposed to left ureteral obstruction (UO), which continued for 24 hours, and then was released. 2 days after release of UO or at the corresponding time point in the non-UO sham-operated mice, the left kidneys were harvested and subjected to immunoblot analyses of HIF-2α and co-detection of TBP as a loading control

    Nuclear extracts were isolated from harvested whole kidneys using NE-PER Nuclear and Cytoplasmic Extraction Reagents supplemented with Complete Protease Inhibitor Cocktail Tablets. Nuclear protein fractions were electrophoresed on 10% SDS-PAGE under reducing conditions and transferred to a nitrocellulose membrane. Membranes were blocked with LI-COR blocking buffer (LI-COR Biosciences, Lincoln, NE). Membranes were then incubated with the same blocking solution containing rabbit polyclonal primary antibodies against HIF-2α (1:500, ab199). After washing, membranes were incubated at room temperature for 1 h in TBS/0.05% Tween 20 buffer with the IRDye800 secondary antibodies (1:10000) and then washed again in TBS/0.05% Tween 20 for 3 times. The blot was visualized using an Odyssey infrared imaging system. All values were normalized to a loading control TATA binding protein (TBP, 1:2000, ab818, Abcam) and expressed as fold increase relative to control.

  • ICC/IF image of ab199 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab199, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-HIF-2-alpha antibody - ChIP Grade (ab199) at 1/500 dilution

    Lane 1 : 20ug of whole cell lysate from PC12 cells.
    Lane 2 : 20ug of whole cell lysate from PC12 cells exposed to hypoxic conditions (12 hours at 3% oxygen).
    Lane 3 : 20ug of whole cell lysate from PC12 cells exposed to hypoxic conditions (12 hours at 3% oxygen) and incubated with antisense HIF2 alpha oligonucleotides 72 hours prior to treatment.

    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 100 kDa
    Observed band size : 96 kDa (why is the actual band size different from the predicted?)

    This image is courtesy of Marilo Dolores Chiara

    The blots were probed with anti-alpha tubulin as a loading control.

Anti-HIF-2-alpha antibody - ChIP Grade (ab199) 使用論文

This product has been referenced in:
  • Bourseau-Guilmain E  et al. Hypoxia regulates global membrane protein endocytosis through caveolin-1 in cancer cells. Nat Commun 7:11371 (2016). WB . Read more (PubMed: 27094744) »
  • Smith SJ  et al. Endothelial-like malignant glioma cells in dynamic three dimensional culture identifies a role for VEGF and FGFR in a tumor-derived angiogenic response. Oncotarget 6:22191-205 (2015). IHC ; Human . Read more (PubMed: 26203665) »

See all 29 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

投稿 Jun 02 2014

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
Username

Dawn Bowles

Verified customer

投稿 Jan 10 2013

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. Fo...

Read More

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, i...

Read More

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 th...

Read More

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, ...

Read More

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 info...

Read More

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 requeste...

Read More

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...

Read More



I am very pleased to hear you would like to accept our offer and test ab199 and ab2185 in ChIP-seq. These codeswill give you 1 free primary antibody eachbefore the expiration date. To redeem this offer, please submit an Abreview forChIP-seq ...

Read More

1-10 of 20 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"