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Setting up a kinase profiler with IMAP

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APPENDIX A
This Application Note illustrates how to efficiently profile kinases using the IMAP assay platform. Appendix A shows in more detail how this Profiler is set up. Please refer to IMAP Application Note #6 for initial substrate selection and enzyme titration curves. All of the Substrate Finder peptides are available from Molecular Devices. (Contact Molecular Devices Customer Service or go to www.moleculardevices.com for details.) Table 4 shows the list of kinases we will profile and includes details of each reaction, substrate sequence and Binding Solution for each substrate. There are 12 kinases assayed utilizing 10 different substrates. The AMPK requires AMP in the reaction for full activation, and the 3 tyrosine kinase reactions contain MnCl2. Binding Solutions are listed as % Buffer A/% Buffer B/dilution of BR. The EC50s determined for each kinase are listed. See IMAP Application Note #6:
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