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Journal of Biomolecular Screening
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What's this?

A Generic High-Throughput Screening Assay for Kinases: Protein Kinase A as an Example

Rommel Mallari

Elissa Swearingen

Wei Liu

Arnold Ow

Stephen W. Young

Shu-Gui Huang

Lead Discovery Department, AGY Therapeutics, Inc., 290 Utah Ave., South San Francisco, CA 94080. fhuang@agyinc.com

A generic high-throughput screening assay based on the scintillation proximity assay technology has been developed for protein kinases. In this assay, the biotinylated 33P-peptide product is captured onto polylysine Ysi bead via avidin. The scintillation signal measuring the product formation increases linearly with avidin concentration due to effective capture of the product on the bead surface via strong coulombic interactions. This novel assay has been optimized and validated in 384-well microplates. In a pilot screen, a signal-to-noise ratio of 5-to 9-fold and a Z' factor ranging from 0.6 to 0.8 were observed, demonstrating the suitability of this assay for high-throughput screening of random chemical libraries for kinase inhibitors. (Journal of Biomolecular Screening 2003:198-204)

Key Words: protein kinase assay • high-throughput screening • scintillation proximity assay • protein kinase A

Journal of Biomolecular Screening, Vol. 8, No. 2, 198-204 (2003)
DOI: 10.1177/1087057103252306
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[Abstract] [PDF]