Recombinant GST-fusion human HIF-1a protein (aa432-528)
HIF is a transcriptional complex that plays a central role in mammalian oxygen homeostasis. Post-translational modification by prolyl hydroxylation is 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 dimerizes 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 are a target for therapeutic manipulation. There are several HIF factors, which include HIF 1-alpha, HIF 1-beta, HIF 2-alpha. HIF 1-alpha ia a 812aa protein in rat and 836aa long in mouse and human (chr 14q21-14q24) with a molecular weight of ~96kD. Under hypoxic conditions, HIF-1a activates the transcription of over 40 genes, many of whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. HIF-1a plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. It is ubiquitous in the cytoplasmic in normoxia, and undergoes nuclear translocation in response to hypoxia. Applications: Suitable for use in ELISA. Western Blot, though not tested, may potentially be used as an application. Other applications not tested. Recommended Dilution: ELISA: 1:10,000-1:30,000 Western Blot: 1:200-1:2000 using ECL. Mammalian cells cultured under reduced (hypoxic) conditions. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 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:
[7C35]
Isotype:
IgG2b
Purity:
Purified by Protein A/G affinity chromatography.
Form:
Supplied as a liquid in PBS, pH 7.4, 0.1% BSA, 40% glycerol.
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