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Setting up a kinase profiler with IMAP

round and maximize the FP signal.

The three substrates that provided the greatest DmP in the assay shown in Figure 17B were then chosen for a subsequent TIE2 titration assay, presented in Figure 18. All of these substrates are part of the MDC peptide inventory. In the SF Tyr plate, their well IDs are as follows: C13D14 (Blk/Lyntide), C7D8 (CSKtide) and K17L18 (PDGFRtide); their sequences and Binding Solutions are also given in Table 3. The TIE2 substrate that ranked the best in terms of DmP and EC50 was the FAM-Blk/Lyntide, so this peptide was chosen for the profiling assay.


SETTING UP THE PROFILER SYSTEM
Now that the specifics of each enzyme assay have been determined, the Profiler can be assembled. 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 updated Profiler from Table 1, including any special reaction requirements, the substrate to use and the optimized binding solution for each substrate.

For kinase selectivity profiling, the screen can be set up in a wide variety of formats. It can be designed
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