Human Phosphotylinosital 3 kinase (PI3K) ELISA Kit

Phosphoinositide 3-kinases (PI3Ks) phosphorylate the 3-prime OH position of the inositol ring of inositol lipids. They have been implicated as participants in signaling pathways regulating cell growth by virtue of their activation in response to various mitogenic stimuli. PI3Ks are composed of a 110-kD catalytic subunit, such as PIK3CB, and an 85-kD adaptor subunit. The predicted 1,070-amino acid protein, called p110-beta by them, is 42% identical to that of bovine p110. Northern blot analysis revealed that the major 4.8-kb p110-beta transcript was expressed in several human and rodent cell lines, as well as in all mouse tissues tested. Minor larger transcripts were detected in some tissues and cell lines.

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Human Phosphotylinosital 3 kinase (PI3K) ELISA Kit has high sensitivity and excellent specificity for detection of Human PI3K. No significant cross-reactivity or interference between Human PI3K and analogues was observed.

This Human Phosphotylinosital 3 kinase (PI3K) ELISA Kit employs a two-site sandwich ELISA to quantitate PI3K in samples. An antibody specific for PI3K has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPI3K present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PI3K is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of PI3K bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.