PI3-Kinase Activity AlphaScreen® Assay

Catalog No.:  K-1300

Description:  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.

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.

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. D'Angelo, 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.

Related Products:
Class I PI3K Activity Assays: Echelon has multiple formats available. Please see the PI3-K Assay Comparison Chart for more information.
Class III PI3K Activity Assay (K-3000)
Lipid Mass ELISA's: PIP3 Mass ELISA (K-2500s) and PI(4,5)P2 Mass ELISA (K-4500)
Human PI3K Enzymes: alpha (p110α/p85α), beta (p110β/p85alphα), PI3Kααα E545K – Active Mutant (p110α/p85α)
Mouse PI3K Enzyme: PI3K (p110α/p85α)
PI3K Inhibitors
Optimized PI3K enzyme buffer (K-KBZ)

Notice 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

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 for details

Options/Sizes Pricing
K-1300-1ea $ 384.00
Additional Information:
Tech Data Sheet - TDS_K-1300_Rev11

PCA - K-1300

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assay and reagents for drug discovery in lipid signaling pathways