Anti-HIF-2-alpha抗体(ab199)
Key features and details
- Rabbit polyclonal to HIF-2-alpha
- Suitable for: ICC/IF, WB
- Reacts with: Rat, Human
- Isotype: IgG
概述
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产品名称
Anti-HIF-2-alpha抗体
参阅全部 HIF-2-alpha 一抗 -
描述
兔多克隆抗体to HIF-2-alpha -
宿主
Rabbit -
特异性
Specific for HIF-2-alpha / EPAS 1. Does not cross react with HIF-1-alpha. -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Rat
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免疫原
Synthetic peptide corresponding to Mouse HIF-2-alpha aa 632-646.
Sequence:GRSNTQWPPDPPLHF
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.40
Preservative: 0.05% Sodium azide
Constituent: 99% PBS -
Concentration information loading... -
纯度
Affinity purified -
克隆
多克隆 -
同种型
IgG -
研究领域
相关产品
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
应用
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| 应用 | Species |
|---|---|
| ICC/IF |
Human
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| WB |
Rat
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| 应用 | Ab评论 | 说明 |
|---|---|---|
| ICC/IF |
Use a concentration of 1 µg/ml.
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| WB | (4) |
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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| 说明 |
|---|
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ICC/IF
Use a concentration of 1 µg/ml. |
|
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!). |
靶标
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功能
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. -
组织特异性
Expressed in most tissues, with highest levels in placenta, lung and heart. Selectively expressed in endothelial cells. -
疾病相关
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. -
序列相似性
Contains 1 basic helix-loop-helix (bHLH) domain.
Contains 1 PAC (PAS-associated C-terminal) domain.
Contains 2 PAS (PER-ARNT-SIM) domains. -
翻译后修饰
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. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- 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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别名
- Basic helix loop helix PAS protein MOP2 antibody
- Basic-helix-loop-helix-PAS protein MOP2 antibody
- bHLHe73 antibody
see all
图片
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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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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.
实验方案
文献 (90)
ab199 被引用在 90 文献中.
- 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
- Luo Y et al. CD146-HIF-1a hypoxic reprogramming drives vascular remodeling and pulmonary arterial hypertension. Nat Commun 10:3551 (2019). PubMed: 31391533
- Wang Y et al. Berberine improves insulin resistance in adipocyte models by regulating the methylation of hypoxia-inducible factor-3a. Biosci Rep 39:N/A (2019). PubMed: 31652442
