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. Featured in Publications 1. Drees, B. E., A. Weipert, et al. (2003). Competitive fluorescence polarization assays for the detection of phosphoinositide kinase and phosphatase activity. Comb Chem High Throughput Screen 6(4): 321-330. 2. Boldyreff, B., T. L. Rasmussen, et al. (2008). Expression and Purification of PI3 Kinase {alpha} and Development of an ATP Depletion and an AlphaScreen PI3 Kinase Activity Assay. J Biomol Screen. 3. DAngelo, N. D., T. S. Kim, et al. (2011). Discovery and optimization of a series of benzothiazole phosphoinositide 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) dual inhibitors. J Med Chem 54(6): 1789-1811. Note to Purchaser Echelon Biosciences products are sold for research and development purposes only and are not to be incorporated into products for resale without written permission from Echelon Biosciences. This kit and all non-radioactive, competitive assays for determining phosphoinositide-3-kinase (PI3-K) activity are covered by Echelon Biosciences Inc. U.S. Patent 7,067,269. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchasers own internal research. For inquiries email busdev echelon-inc.com. Product Keywords: PI3-Kinase, PI3K, Phosphatidylinositol 3-kinases, PI 3-kinases, PI3Ks, K-1000, PIP3, PI(345)P3, PtdIns(3,4,5)P3, Phosphatidylinositol (3,4,5)-triphosphate Bulk Discounts: Bulk discounts are available, please email echelon echelon-inc.com for details
Documents Technical Data Sheet PI3K Assay Comparison Guide |
