製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to HIF3 alpha
  • 特異性
    We have observed both nuclear and cytoplasmic localisation in IF with this antibody and we believe this could be due to dysregulated expression of HIF3 alpha in our cell lines.
  • アプリケーション
    適用あり: WB, ICC/IF, IP, IHC-Pmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 300 - 400 of Human HIF3 alpha.

    (Peptide available as ab20077.)

  • ポジティブ・コントロール
    • This antibody gives a positive signal in human heart tissue lysate.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab10134 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
WB 1/500. Detects a band of approximately 60 kDa (predicted molecular weight: 72 kDa).Can be blocked with Human HIF3 alpha peptide (ab20077).
ICC/IF Use a concentration of 1 - 5 µg/ml.
IP Use at an assay dependent concentration.
IHC-P 1/100.

ターゲット情報

  • 機能
    Involved in adaptive response to hypoxia. Suppresses hypoxia-inducible expression of HIF1A and EPAS1. Binds to core DNA sequence 5'-TACGTG-3' within the hypoxia response element (HRE) of target gene promoters. The complex HIF3A-ARNT activates the transcription of reporter genes driven by HRE. Isoform 4 has a dominant-negative function of inactivating HIF1A-mediated transcription. Isoform 4 attenuates the binding of HIF1A to hypoxia-responsive elements (HRE), thus inhibiting HRE-driven transcription. Hypoxia induces down-regulation of isoform 4, leading to activation of HIF1A in hypoxia. Conversely, upon restoring normoxia, the expression of isoform 4 increases and thereby secure an inhibition of HIF1A activity. Isoform 4 may be a negative regulator of hypoxia-inducible gene expression in the kidney and may be involved in renal tumorigenesis. Functions as an inhibitor of angiogenesis in the cornea.
  • 組織特異性
    Expressed in kidney. Expressed abundantly in lung epithelial cells. Expression is regulated in an oxygen-dependent manner.
  • 配列類似性
    Contains 1 basic helix-loop-helix (bHLH) domain.
    Contains 2 PAS (PER-ARNT-SIM) domains.
  • 翻訳後修飾
    In normoxia, hydroxylated on Pro-492 in the oxygen-dependent degradation domain (ODD) by PHD. The hydroxylated proline promotes interaction with VHL, initiating rapid ubiquitination and subsequent proteasomal degradation.
  • 細胞内局在
    Nucleus. Cytoplasm. In the nuclei of all periportal and perivenous hepatocytes. In the distal perivenous zone, detected in the cytoplasm of the hepatocytes.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Basic-helix-loop-helix-PAS protein MOP7 antibody
    • bHLHe17 antibody
    • Class E basic helix-loop-helix protein 17 antibody
    • HIF 3A antibody
    • HIF 3A4 antibody
    • HIF-3-alpha antibody
    • HIF3 alpha antibody
    • HIF3-alpha antibody
    • HIF3-alpha-1 antibody
    • HIF3A antibody
    • HIF3A_HUMAN antibody
    • Hypoxia Inducible Factor 3 alpha antibody
    • Hypoxia inducible factor 3 alpha subunit antibody
    • Hypoxia inducible factor three alpha antibody
    • Hypoxia-inducible factor 3-alpha antibody
    • Inhibitory PAS domain protein antibody
    • IPAS antibody
    • Member of PAS protein 7 antibody
    • MOP7 antibody
    • PAS domain-containing protein 7 antibody
    • PASD7 antibody
    see all

画像

  • ICC/IF image of ab10134 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab10134, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • ICC/IF image of ab10134 stained HeLa 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 (ab10134, 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.
  • Anti-HIF3 alpha antibody (ab10134) at 2 µg/ml + Human heart tissue lysate - total protein (ab29431) at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 72 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)


    Exposure time : 8 minutes

参考文献

This product has been referenced in:
  • Rao K  et al. Protective effect of zinc preconditioning against renal ischemia reperfusion injury is dose dependent. PLoS One 12:e0180028 (2017). WB ; Human . Read more (PubMed: 28686686) »
  • Fala AM  et al. Unsaturated fatty acids as high-affinity ligands of the C-terminal Per-ARNT-Sim domain from the Hypoxia-inducible factor 3a. Sci Rep 5:12698 (2015). Read more (PubMed: 26237540) »

See all 8 Publications for this product

レビューと Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (small bowel adenoma)
Specification
small bowel adenoma
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Jul 31 2007

Thank you for your enquiry. Do you know the name of the person who does the purchasing from your lab? I will only be able to give you a replacement for the antibody. We can only replace or refund the product that was defective, typically only with...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (in vitro translated HIF-3a1)
Specification
in vitro translated HIF-3a1
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4%
Username

Abcam user community

Verified customer

投稿 Feb 22 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - other (IVT HIF-3a1)
Specification
IVT HIF-3a1
Immuno-precipitation step
Protein A
Username

Abcam user community

Verified customer

投稿 Feb 07 2006

Thank you for your enquiry. I blasted the immunogen sequence and found that the sequence is identical in isoforms a, b, c, and d. I was unable to find the other two splice variants. If you could send me the sequences of the other two isoforms, I c...

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Thank you very much for providing the details of your protocol. I would like to try RIPA buffer which contains the ionic detergent sodium deoxycholate as an active constituent and is particularly used for nuclear membrane disruption for nuclear ext...

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Thank you for your email and I'm sorry to hear that you are experiencing difficulty with ab10134. Could we get a detailed protocol from you, please? We have some general questions, the answers to which will enable us to investigate this matter as quick...

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

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