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Setting up a kinase profiler with IMAP

IMAP APPLICATION NOTE #6

By Liz Gaudet, Ph.D., Joyce Itatani, Francisco Ramirez and Annegret Boge, Ph.D. Molecular Devices Corporation, 1311 Orleans Dr., Sunnyvale, CA


INTRODUCTION
Kinase profiling is a key aspect of lead compound drug discovery. Many suppliers offer a profiling service, but for maximum flexibility as well as reasons of confidentiality, many researchers would prefer to do profiling themselves instead of sending their compounds to another company. IMAP is perfect for this task because it adapts easily from its major role in high-throughput screening to the demands of kinase profiling.

This application note describes how to set up and run kinase profiling (Profiler) experiments using IMAP. The IMAP fluorescence polarization (FP) assay platform is applicable to a variety of enzymes, especially protein kinases. IMAP is based on the high affinity binding of phosphate to immobilized trivalent metals, rather than on antibody recognition. Thus virtually any peptide substrate can be easily configured to run in an IMAP assay.

This generic feature allows IMAP to be used to test multiple kinases efficiently using IMAP Substrate Finder kits and the IMAP Progressive Binding System. These kinases then become the panel for a Profiler that runs efficiently and precisely in IMAP. You can rapidly test many compounds in the Profiler to assess selectivity, determine IC50s, and assay with a wide range (up
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