CST, 3434T, Hydroxy-HIF-1 alpha (Pro564) (D43B5) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying HIF1A (Pro564) hydroxylate. Validated for Western Blotting,Immunoprecipitation,Immunofluorescence (Immunocytochemistry). Available in 2 sizes. Highly specific and rigorously validated in-house, Hydroxy-HIF-1 alpha (Pro564) (D43B5) Rabbit Monoclonal Antibody (CST #3434) is ready to ship. Product Usage Information Western Blotting: 1:1000 Immunoprecipitation: 1:50 Immunofluorescence (Immunocytochemistry): 1:3200 - 1:6400 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunofluorescence (Immunocytochemistry) Specificity / Sensitivity Hydroxy-HIF-1 alpha (Pro564) (D43B5) Rabbit Monoclonal Antibody detects endogenous levels of HIF-1α only when hydroxylated at Pro564. This antibody may cross react with other overexpressed proline hydroxylated proteins. Species Reactivity: Human, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic hydroxypeptide corresponding to residues surrounding Pro564 of human HIF-1α. Background 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 (VHL) 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 xenobiotic 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). Alternate Names ARNT interacting protein; ARNT-interacting protein; Basic-helix-loop-helix-PAS protein MOP1; BHLHE78; Class E basic helix-loop-helix protein 78; HIF-1-alpha; HIF-1A; HIF-1alpha; HIF1; HIF1-alpha; HIF1A; hypoxia inducible factor 1 alpha subunit; hypoxia inducible factor 1 subunit alpha; hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor); hypoxia-inducible factor 1 alpha isoform I.3; Hypoxia-inducible factor 1-alpha; hypoxia-inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor); hypoxia-inducible factor1alpha; Member of PAS protein 1; member of PAS superfamily 1; MOP1; PAS domain-containing protein 8; PASD8 Specification REACTIVITY: H Mk SENSITIVITY: Endogenous MW (kDa): 120 Source/Isotype: Rabbit IgG