Product nameAnti-HIF3 alpha/IPAS antibody
See all HIF3 alpha/IPAS primary antibodies
DescriptionRabbit polyclonal to HIF3 alpha/IPAS
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 applicationsSuitable for: WB, ICC/IF, IP, IHC-Pmore details
Species reactivityReacts with: Human
Synthetic peptide corresponding to Human HIF3 alpha/IPAS aa 350-450 conjugated to keyhole limpet haemocyanin.
(Peptide available as
- This antibody gives a positive signal in human heart tissue lysate.
Previously labelled as HIF3 alpha.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
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.
|WB||1/500. Detects a band of approximately 60 kDa (predicted molecular weight: 72 kDa).Can be blocked with Human HIF3 alpha/IPAS peptide (ab20077).|
|ICC/IF||Use a concentration of 1 - 5 µg/ml.|
|IP||Use at an assay dependent concentration.|
FunctionInvolved 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 specificityExpressed in kidney. Expressed abundantly in lung epithelial cells. Expression is regulated in an oxygen-dependent manner.
Sequence similaritiesContains 1 basic helix-loop-helix (bHLH) domain.
Contains 2 PAS (PER-ARNT-SIM) domains.
modificationsIn 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 localizationNucleus. 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
- Basic-helix-loop-helix-PAS protein MOP7 antibody
- bHLHe17 antibody
- Class E basic helix-loop-helix protein 17 antibody
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/IPAS antibody (ab10134) at 2 µg/ml + Human heart tissue lysate - total protein (ab29431) at 20 µg
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.
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) »