Anti-HIF-2-alpha 抗体 (ab199)
Key features and details
- Rabbit polyclonal to HIF-2-alpha
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Rat, Human, African green monkey
- Isotype: IgG
製品の概要
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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: ICC/IF, IHC-P, WBmore details -
種交差性
交差種: Rat, Human, African green monkey -
免疫原
Synthetic peptide corresponding to Mouse HIF-2-alpha aa 632-646.
Sequence:GRSNTQWPPDPPLHF
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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
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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
関連製品
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Compatible Secondaries
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Conjugation kits
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Isotype control
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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 |
|---|---|---|
| ICC/IF |
Use a concentration of 1 µg/ml.
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| IHC-P | (1) |
1/100.
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| 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!).
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| 特記事項 |
|---|
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ICC/IF
Use a concentration of 1 µg/ml. |
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IHC-P
1/100. |
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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
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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
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参照データベース
- Entrez Gene: 2034 Human
- Entrez Gene: 29452 Rat
- Omim: 603349 Human
- SwissProt: Q99814 Human
- SwissProt: Q9JHS1 Rat
- Unigene: 468410 Human
- Unigene: 55138 Rat
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別名
- Basic helix loop helix PAS protein MOP2 antibody
- Basic-helix-loop-helix-PAS protein MOP2 antibody
- bHLHe73 antibody
see all
画像
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Western blot - Anti-HIF-2-alpha antibody (ab199)
Analysis on normoxic and hypoxic nuclear rat cell lysates.
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Western blot - Anti-HIF-2-alpha antibody (ab199)Image from PLoS One. 2012; 7(1): e29876. Fig3A, doi: 10.1371/journal.pone.0029876 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/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.
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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.
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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 -
Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody (ab199)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.
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Western blot - Anti-HIF-2-alpha antibody (ab199)This image is courtesy of Marilo Dolores ChiaraAll lanes : Anti-HIF-2-alpha antibody (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?
The blots were probed with anti-alpha tubulin as a loading control.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (95)
ab199 は 95 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