HIF-1 Alpha Cell Based (HIF) 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. 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: hydroxylation of two proline residues and acetylation of a lysine residue in its oxygen dependent degradation domain (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 Cell Based ELISA Kit is an immunoassay developed for rapid detection of HIF-1 Alpha in fixed cells. Cells on a microplate are stimulated for HIF-1 Alpha stabilization, fixed, permeabilized, and then neutralized in the well. HIF-1 Alpha is then detected with an 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.

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Small volumes of HIF-1 Alpha Cell Based elisa kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS168874 is a ready-to-use microwell, strip-or-full plate ELISA (enzyme-linked immunosorbent assay) Kit for analyzing the presence of the HIF-1 Alpha Cell Based, ELISA Kit target analytes in biological samples. The concentration gradients of the kit standards or positive controls render a theoretical kit detection range in biological research samples containing HIF-1 Alpha Cell Based. The ELISA analytical biochemical technique of the MBS168874 kit is based on HIF-1 Alpha Cell Based antibody-HIF-1 Alpha Cell Based antigen interactions (immunosorbency) and an HRP colorimetric detection system to detect HIF-1 Alpha Cell Based antigen targets in samples. The ELISA Kit is designed to detect native, not recombinant, HIF-1 Alpha Cell Based. Appropriate sample types may include undiluted body fluids and/or tissue homogenates, secretions. Quality control assays assessing reproducibility identified the intra-assay CV (%) and inter-assay CV(%).