The PI3-Kinase Activity AlphaScreen® Assay is a sensitive and robust method for the quantification of PI(3,4,5)P3 (PIP3), the end product of Class I PI3K reactions. This high-throughput (384-well) assay can be used for in vitro determination of PI3K activity in cancer tissues and cell lines and compound screening for drug discovery.

Assay Range:0.46 NM to 1,000 nM

Sample Type:purified enzyme, inhibitor screen

Sample Volume:2.5 µL PI3-K inhibitor

Product Background:When PIP3 is generated via phosphorylation of PI(4,5)P2 (PIP2) by PI3-Kinase, the products of the enzymatic reaction compete with biotinylated I(1,3,4,5)P4 for the interaction with the PIP3 detector protein. The interaction of biotinylated-I(1,3,4,5)P4 and the PIP3 detector protein brings both Acceptor and Donor beads together, producing a cascade of chemical reactions and leading to the AlphaScreen® signal. In the absence of this interaction, proximity of the Donor and Acceptor beads is decreased, producing a loss of luminescent signal which is inversely related to enzyme activity. AlphaScreen® technology is based on the emission of light (520-620nm) by Acceptor beads activated by the proximity of Donor beads.

Echelon has multiple formats available for Class III PI3-K. Please see the PI3-K Assay Comparison Guide below for more information.

Things to consider:This product is intended for use with the Amplified Luminescent Proximity Homogenous Assay (AlphaScreen®) technology and requires the researcher to obtain donor and acceptor beads from the AlphaScreen® GST (Glutathione-S-Transferase) Detection Kit, which must be purchased separately from PerkinElmer Life Sciences. A microplate reader with AlphaScreen® capabilities is also required.