Recombinant human purified HIF-1 alpha protein (aa530-826, GST fusion protein)
HIF is a transcriptional complex that plays a central role in mammalian oxygen homeostasis, the posttranslational modification by prolyl hydroxylation as a key regulatory event that targets HIF-alpha subunits for proteasomal destruction via the von Hippel-Lindau ubiquitylation complex. The transcriptional complex is composed of an alpha-beta heterodimer, HIF-beta being a constitutive nuclear protein that dimerises with oxygen regulated HIF-alpha subunits. In normoxia, 4-hydroxylation of human HIF-alpha at Pro402 or Pro564 by a set of HIF prolyl hydroxylase isoenzymes (PHD 1-3) mediates HIF1-alpha recognition by von Hippel-Lindau ubiquitin ligase complex leading to its proteasomal destruction. In hypoxia (deprivation of oxygen), lack of hydroxylase activity enables HIF-alpha subunits to escape destruction and become transcriptionally active. Thus HIF hydroxylases provide a focus for understanding cellular responses to hypoxia and target for therapeutic manipulation. There are several HIF factors, which include HIF 1-alpha, HIF 1-beta, HIF 2-alpha HIF 1-alpha: A 812aa protein in rat and 836aa long in mouse and human (chr 14q21-14q24) Mol.wt of ~96kD. A master transcriptional regulator of the adaptive response to hypoxia. Under hypoxic conditions activates the transcription of over 40 genes, including, erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. Plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. It is ubiquitous in expression as cytoplasmic in normoxia, nuclear translocation in response to hypoxia. Applications: Suitable for use in Western Blot and ELISA. Other applications not tested. Recommended Dilution: Western Blot: 1:200-1:2000 using Chemiluminescence technique. Detected ~120kD bands representing post translationally modified forms of HIF-1a. Mammalian cells cultured under reduced (hypoxic) conditions. ELISA: 1:10,000-1:30,000 Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder may be stored at -20C. Stable for 12 months at -20C. Reconstitute with sterile ddH2O or PBS. Aliquot to avoid repeated freezing and thawing. Store at -20C. Reconstituted product is stable for 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Clonality:
Monoclonal
Clone Designation:
[7C37]
Isotype:
IgG1
Purity:
Purified by immunoaffinity chromatography.
Form:
Supplied as a lyophilized powder from PBS, 0.05% sodium azide.
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