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Cardiovascular Hypoxia HIF

Anti-HIF-2-alpha抗体(ab199)

  • Datasheet
  • SDS
Reviews (6)Q&A (15)References (90)

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Western blot - Anti-HIF-2-alpha antibody (ab199)
  • Western blot - Anti-HIF-2-alpha antibody (ab199)
  • Immunocytochemistry/ Immunofluorescence - 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: ICC/IF, WB
  • Reacts with: Rat, Human
  • Isotype: IgG

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概述

  • 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

    Application Species
    ICC/IF
    Human
    WB
    Rat
    See all applications and species data
  • 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

性能

  • 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

    • 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

相关产品

  • 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)

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
ICC/IF
Human
WB
Rat
应用 Ab评论 说明
ICC/IF
Use a concentration of 1 µg/ml.
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!).
说明
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

  • 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.
  • 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
  • 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
    • 别名

      • 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

    图片

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

      Analysis on normoxic and hypoxic nuclear rat cell lysates.

    • Western blot - Anti-HIF-2-alpha antibody (ab199)
      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody (ab199)
      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.

    • Western blot - Anti-HIF-2-alpha antibody (ab199)
      Western blot - Anti-HIF-2-alpha antibody (ab199)This image is courtesy of Marilo Dolores Chiara
      All 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.

      See Abreview

    实验方案

    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • Datasheet
    • SDS
  • 文献 (90)

    发表研究结果有使用 ab199?请让我们知道,以便我们可以引用本数据表中的参考文章。

    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
    View all Publications for this product

    客户评价及客户问答

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    提交评价 提交问题

    1-4 of 4 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

    提交于 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

    提交于 Jan 10 2013

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HUVEC)
    Loading amount
    20 µg
    Specification
    HUVEC
    Treatment
    Hypoxia 4 h
    Gel Running Conditions
    Reduced Denaturing (10 %)
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    提交于 Mar 22 2011

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293 cells transfected with human HIF-2alpha expres)
    Loading amount
    100000 cells
    Specification
    293 cells transfected with human HIF-2alpha expres
    Treatment
    Cells were incubated in hypoxic condition (1% Oxygen) for 24hrs
    Gel Running Conditions
    Non-reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Sep 04 2009

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