Product Pathways - Protein Translation
Hydroxy-HIF-1α (Pro564) (D43B5) XP® Rabbit mAb #3434
|3434S||100 µl (10 western blots)||---||In Stock||---|
|3434P||40 µl (4 western blots)||---||In Stock||---|
|3434||carrier free and custom formulation / quantity||email request|
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Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry)
Species predicted to react based on 100% sequence homology: Mouse, Rat, Monkey, Chicken, Xenopus, Zebrafish, Pig.
Specificity / Sensitivity
Hydroxy-HIF-1α (Pro564) (D43B5) XP® Rabbit mAb detects endogenous levels of HIF-1α only when hydroxylated at Pro564. This antibody may cross react with other overexpressed proline hydroxylated proteins.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic hydroxypeptide corresponding to residues surrounding Pro564 of human HIF-1α.
Western blot analysis of extracts from HeLa cells, treated with either 10 μM of MG132 (to accumulate hydroxylated HIF-1α) or 10 µM MG132 and 1 mM DMOG (to accumulate nonhyroxylated HIF-1α), using Hydroxy-HIF-1α (Pro564) (D43B5) XP® Rabbit mAb (upper) or total HIF-1α Antibody #3716 (lower).
Hypoxia-inducible factor 1 (HIF1) is a heterodimeric transcription factor that plays a critical role in the cellular response to hypoxia (1). The HIF1 complex consists of two subunits, HIF-1α and HIF-1β, which are basic helix-loop-helix proteins of the PAS (Per, ARNT, Sim) family (2). HIF1 regulates the transcription of a broad range of genes that facilitate responses to the hypoxic environment, including genes regulating angiogenesis, erythropoiesis, cell cycle, metabolism, and apoptosis. The widely expressed HIF-1α is typically degraded rapidly in normoxic cells by the ubiquitin/proteasomal pathway. Under normoxic conditions, HIF-1α is proline hydroxylated leading to a conformational change that promotes binding to the von Hippel Lindau protein (VLH) E3 ligase complex; ubiquitination and proteasomal degradation follows (3,4). Both hypoxic conditions and chemical hydroxylase inhibitors (such as desferrioxamine and cobalt) inhibit HIF-1α degradation and lead to its stabilization. In addition, HIF-1α can be induced in an oxygen-independent manner by various cytokines through the PI3K-AKT-mTOR pathway (5-7).
HIF-1β is also known as AhR nuclear translocator (ARNT) due to its ability to partner with the aryl hydrocarbon receptor (AhR) to form a heterodimeric transcription factor complex (8). Together with AhR, HIF-1β plays an important role in xenobiotics metabolism (8). In addition, a chromosomal translocation leading to a TEL-ARNT fusion protein is associated with acute myeloblastic leukemia (9). Studies also found that ARNT/HIF-1β expression levels decrease significantly in pancreatic islets from patients with type 2 diabetes, suggesting that HIF-1β plays an important role in pancreatic β-cell function (10).
Two critical prolines in HIF-1α (Pro564 and Pro402) can be hydroxylated by proline hydroxylase under normoxia conditions. Hydroxylation of HIF-1α leads to its binding to VHL and ubiquitin mediated degradation (3,11,12).
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- Jiang, B.H. et al. (2001) Cell Growth Differ 12, 363-9.
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- Walisser, J.A. et al. (2004) Proc Natl Acad Sci U S A 101, 16677-82.
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- Gunton, J.E. et al. (2005) Cell 122, 337-49.
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- Masson, N. et al. (2001) EMBO J 20, 5197-206.
- Tian, Y.M. et al. (2011) J Biol Chem 286, 13041-51. Applications: Western Blotting.
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For Research Use Only. Not For Use In Diagnostic Procedures.
XP® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.