Hypoxia Inducible Factor 1a (HIF-1a)

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.\n\nApplications:\nSuitable for use in ELISA and Western Blot. Other applications not tested. \n\nRecommended Dilution:\nELISA: 1:10,000-1:30,000\nWestern Blot: 1:200-1:2000 using ECL. Mammalian cells cultured under reduced (hypoxic) conditions.\nOptimal dilutions to be determined by the researcher. \n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. 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.

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SPECIFICATIONS

Catalog Number

H9807-06

Size

50ug

Applications

ELISA, WB

Hosts

Mouse

Reactivities

Hum, Mouse, Rat, Sheep

Form

Supplied as a liquid in PBS, pH 7.4, 0.1% BSA, 40% glycerol.

P Type

Mab

Purity

Purified by Protein A/G affinity chromatography.

Isotype

IgG2b

References

1. Kaelin, W.G., et al., Nature 399: 203 (1999). 2. Iyer, N.V., et al., Genes and Dev. 12: 149 (1998). 3. Semenza, G.L., et al., J. Lab. Clin. Med. 131: 207 (1998). 4. Nguyen, S.V., et al., Biochem. Biophys. Res. Comm. 265: 382 (1999). 5. Feldser, D., et al., Cancer Res. 59: 3915 (1999).

Additional Info

Recognizes human Hypoxia Inducible Factor 1a. Detects several bands at ~120kD representing post-translationally modified forms of HIF-1a. Species Crossreactivity: Mouse, rat, sheep and ferret.