HIF-1 Alpha ELISA Kit

HIF-1 Alpha Sandwich ELISA Kit

Cat. Num. AAA169099

• Synonyms: HIF-1 Alpha; HIF-1 Alpha Sandwich ELISA Kit; HIF-1 Alpha elisa kit

• Assay Type: Sandwich
• Preparation and Storage: Upon receipt, store the HIF-1 Alpha Standard at -80 degree C and the Anti-HIF-1 Alpha Antibody at -20 degree C. Store all other components at 4 degree C.

Typical Testing Data/Standard Curve (for reference only)

Related Product Information for HIF-1 Alpha elisa kit

Background: Mammalian cells are able to sense low oxygen conditions and turn on a series of genes in response to the lack of oxygen. The hypoxia-inducible factor 1 transcriptional activator complex (HIF-1) is involved in the activation of several hypoxia-responsive genes including erythropoietin and VEGF. The HIF-1 complex is composed of two protein subunits: the constitutively expressed HIF-1b/ARNT (aryl hydrocarbon receptor nuclear translocator), and HIF-1 Alpha, the latter of which is not detected during normoxia since it is continually degraded by the ubiquitin proteasome system (UPS). In the presence of low oxygen conditions, however, HIF-1 Alpha is stabilized, accumulates, translocates from the cytosol to the nucleus, dimerizes with HIF-1b/ARNT, and becomes transcriptionally active. The activated HIF-1 complex then associates with hypoxic response elements (HREs) and binds transcriptional coactivators to induce gene expression. Tight regulation of the stability and function of HIF-1 is controlled by its post-translational modifications, such as hydroxylation, ubiquitination, acetylation, and phosphorylation. Under normal oxygen conditions, the post-translational modification of HIF-1 Alpha occurs within several domains such as the N-terminal transactivation domain, C-terminal transactivation domain, and oxygen dependent degradation domain (ODDD). Hydroxylation of two proline residues and acetylation of a lysine residue in its ODDD promote binding of HIF-1 with the von Hippel-Lindau (pVHL) ubiquitin E3 ligase complex. This pVHL complex modifies HIF-1 with ubiquitin, marking it for degradation by the 26S proteasome. Furthermore, hydroxylation of C-terminal asparagine residue in the c-terminal transactivation domain blocks association of HIF-1 with CBP/p300 and as a result inhibits HIF-1 transcriptional activity. Upon synthesis of HIF-1 Alpha, the protein is rapidly hydroxylated by a family of 2-oxoglutarate dioxygenases on proline 402 and 564. Hypoxic or chemical inactivation of these dioxygenases (which were later termed proline hydroxylase domains (PHDs)), leads to an increase in the half life of HIF-1 Alpha and subsequent activation of HIF-1 complex. HIF-1 Alpha Sandwich ELISA Kit is an immunoassay developed for rapid detection of HIF-1 Alpha in any protein sample. HIF-1 Alpha protein is captured by a mouse monoclonal antibody that is attached to the well. HIF-1 Alpha is then detected with a goat polyclonal anti-HIF-1 Alpha antibody followed by an HRP conjugated secondary antibody. Each kit provides sufficient reagents to perform up to a total of 96 assays, and can detect HIF-1 Alpha from human, mouse, or rat.

Product Categories/Family for HIF-1 Alpha elisa kit

Cell Based Assays; Cell Health Assays >Hypoxia Assays

References

1. Ke Q and Costa M. (2006) Molecular Pharm. 70:1469-1480.
2. Salceda S and Caro J (1997) J. Biol. Chem. 272: 22642-22647.
3. CW Pugh, JF O'Rourke, M Nagao, JM Gleadle, and PJ Ratcliffe (1997) J. Biol. Chem. 272: 11205-11214.
4. LE Huang, Z Arany, DM Livingston, and HF Bunn (1996) J. Biol. Chem. 272: 32253-32259.
5. PJ Kallio, I Pongratz, K Gradin, J McGuire, and L Poellinger (1997) Proc Natl Acad Sci USA 94: 5667-5672.