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Setting up a kinase profiler with IMAP

230 151 40/60/ 1:1500 73 229 0/100/ 1:1500

Note to Table 2: The SF Tyr kit assay well ID, peptide sequence, and part number /name of peptide are shown. The Binding Solutions for the titration assay and for the SF Tyr assays are also shown. Binding Solutions are listed as % Buffer A/% Buffer B/dilution of Binding Reagent. For the FAK titration assay, the Binding Solutions were optimized slightly from the SF Tyr protocol in order to reduce the FP background and maximize the FP signal.

Serial dilutions of FAK were then tested with the three top substrates ranked by DmP from Figure 6, Panel B. Results of the enzyme titration assay are shown in Figure 7, and Table 2 lists the peptide sequences and their names and well IDs. Two of the top peptide hits with 0.02 U/mL are presently peptides in MDC's inventory; these are the FAM-Srctide and FAM-Src-Family-tide (well IDs A11B12 and G13H14, respectively). As expected the SF Tyr assay with the lesser FAK concentration was a more accurate predictor of EC50 than the 0.07 U/mL assay. The FAK substrate that ranked the best in terms of '"/>

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