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Anti-HIF3 alpha/IPAS antibody (ab10134)

Key features and details

  • Rabbit polyclonal to HIF3 alpha/IPAS
  • Suitable for: WB, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-HIF3 alpha/IPAS antibody
    See all HIF3 alpha/IPAS primary antibodies

  • Description

    Rabbit polyclonal to HIF3 alpha/IPAS

  • Host species

    Rabbit

  • Specificity

    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/IPAS in our cell lines.

  • Tested applications

    Suitable for: WB, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide corresponding to Human HIF3 alpha/IPAS aa 350-450 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab20077)

  • Positive control

    • This antibody gives a positive signal in human heart tissue lysate.

  • General notes

    Previously labelled as HIF3 alpha. 

Properties

Applications

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.

Application Abreviews Notes
WB 1/500. Detects a band of approximately 60 kDa (predicted molecular weight: 72 kDa).
ICC/IF Use a concentration of 1 - 5 µg/ml.

Target

  • Function

    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.

  • Tissue specificity

    Expressed in kidney. Expressed abundantly in lung epithelial cells. Expression is regulated in an oxygen-dependent manner.

  • Sequence similarities

    Contains 1 basic helix-loop-helix (bHLH) domain.
    Contains 2 PAS (PER-ARNT-SIM) domains.

  • Post-translational
    modifications

    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.

  • Cellular localization

    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

  • Database links

    • Alternative names

      • 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

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-HIF3 alpha/IPAS antibody (ab10134)
      Immunocytochemistry/ Immunofluorescence - Anti-HIF3 alpha/IPAS antibody (ab10134)

      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).

    • Immunocytochemistry/ Immunofluorescence - Anti-HIF3 alpha/IPAS antibody (ab10134)
      Immunocytochemistry/ Immunofluorescence - Anti-HIF3 alpha/IPAS antibody (ab10134)
      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.
    • Western blot - Anti-HIF3 alpha/IPAS antibody (ab10134)
      Western blot - Anti-HIF3 alpha/IPAS antibody (ab10134)
      Anti-HIF3 alpha/IPAS 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


      This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% milk before being incubated with ab10134 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.  The band detected by ab10134 migrates at approximately 60-kDa.  Although the predicted molecular weight of this protein is 72-kDa, the 60-kDa band has been observed in the literature.  The intensity of this band has been shown to increase in cells exposed to hypoxic conditions (PMID:16775626). 

      Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

    References (9)

    Publishing research using ab10134? Please let us know so that we can cite the reference in this datasheet.

    ab10134 has been referenced in 9 publications.

    • Xu Z  et al. HAND2-AS1 Inhibits Gastric Adenocarcinoma Cells Proliferation and Aerobic Glycolysis via miRNAs Sponge. Cancer Manag Res 12:3053-3068 (2020). PubMed: 32431548
    • Rao K  et al. Protective effect of zinc preconditioning against renal ischemia reperfusion injury is dose dependent. PLoS One 12:e0180028 (2017). WB ; Human . 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). PubMed: 26237540
    • Gong H  et al. HIF2a signaling inhibits adherens junctional disruption in acute lung injury. J Clin Invest 125:652-64 (2015). PubMed: 25574837
    • Gits CM  et al. MicroRNA response to hypoxic stress in soft tissue sarcoma cells: microRNA mediated regulation of HIF3a. BMC Cancer 14:429 (2014). WB ; Human . PubMed: 24927770
    • Che Y  et al. Co-expression of XIAP and cyclin D1 complex correlates with a poor prognosis in patients with hepatocellular carcinoma. Am J Pathol 180:1798-807 (2012). Human . PubMed: 22429965
    • Tanaka T  et al. The human HIF (hypoxia-inducible factor)-3alpha gene is a HIF-1 target gene and may modulate hypoxic gene induction. Biochem J 424:143-51 (2009). PubMed: 19694616
    • Elbarghati L  et al. Effects of hypoxia on transcription factor expression in human monocytes and macrophages. Immunobiology 213:899-908 (2008). WB ; Human . PubMed: 18926304
    • Li QF  et al. Hypoxia upregulates hypoxia inducible factor (HIF)-3alpha expression in lung epithelial cells: characterization and comparison with HIF-1alpha. Cell Res 16:548-58 (2006). PubMed: 16775626

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-HIF3 alpha/IPAS antibody (ab10134)
      Immunocytochemistry/ Immunofluorescence - Anti-HIF3 alpha/IPAS antibody (ab10134)

      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).

    • Immunocytochemistry/ Immunofluorescence - Anti-HIF3 alpha/IPAS antibody (ab10134)
      Immunocytochemistry/ Immunofluorescence - Anti-HIF3 alpha/IPAS antibody (ab10134)
      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.
    • Western blot - Anti-HIF3 alpha/IPAS antibody (ab10134)
      Western blot - Anti-HIF3 alpha/IPAS antibody (ab10134)
      Anti-HIF3 alpha/IPAS 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


      This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% milk before being incubated with ab10134 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.  The band detected by ab10134 migrates at approximately 60-kDa.  Although the predicted molecular weight of this protein is 72-kDa, the 60-kDa band has been observed in the literature.  The intensity of this band has been shown to increase in cells exposed to hypoxic conditions (PMID:16775626). 

      Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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