The 5′ PtdIns(3,4,5)P3 Phosphatase Activity Fluorescence Polarization Assay determines activity through quantification of the end product, PI(3,4)P2.

Sample Type: in vitro determination of 5′ phosphatase activity in cancer tissues and cell lines and compound screening for drug discoverySample Volume: 2.5-5.0 µL enzyme / inhibitorAssay Range: 0.0625 µM to 2.0 µM

Product Background: This 384 well, fluorescence polarization-based, competitive assay measures the phosphatase activity of SHIP2 which hydrolyzes the 5-phosphate from PtdInd(3,4,5)P₃. After the phosphatase reactions are complete, reaction products are mixed with a PI(3,4)P₂ detector protein and the fluorescent PI(3,4)P₂ probe. Polarization (mP) values decrease as probe binding to the PI(3,4)P₂ detector is displaced by PI(3,4)P₂ produced by enzymatic activity and the amount of unbound fluorescent probe in the mixture increases.

Product Keywords: SHIP2, SHIP1, PI(3,4)P2

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 purchaser’s own internal research. For inquiries email busdev@echelon-inc.com.