Commonly used methods for kinase screening either reply on antibody detection of a phospho-epitope, or monitoring ATP depletion as the result of kinase activity in vivo. We have developed an improved, generic kinase assay format that monitors the accumulation of ADP as the result of kinase activity. This approach was initially developed as a very rapid, kinetic application for assay development and compound profiling and characterization. However, the assay formulation has recently been optimized to provide features more suitable for primary screening applications. The benefits of this approach include gain-of-signal output, high DTT tolerance, high ATP tolerance and the ability to use whole protein substrates. The detection of ADP is based on an enzyme-coupled reaction converting ADP to a fluorescent signal. This has traditionally been challenging with issues such as compound interference and overall performance. We will present data demonstrating assay improvements and performance with respect to compound screening.
Life Sciences 2007 (2007) Proc Life Sciences, PC323
Poster Communications: ADP Hunter™ Plus: an optimized assay format with minimized compound interference for kinase primary screening using ADP accumulation
A. Fowler1, N. W. Charter2, W. Feng2, R. Singh2, G. Perez2, K. Olson2, S. Kumar1
1. Discoverx, Birmingham, United Kingdom. 2. Discoverx, Fremont, CA, USA.
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