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Setting up a kinase profiler with IMAP

mbol;mso-symbol-font-family:Symbol'>x 5FAM-ERMRPRKRQGSVRRRV-NH2 19 11 Ros 5FAM-KKKSPGEYVNIEFG-NH2 20 12 TIE2 untested 21

Note to Table 1: Many Molecular Devices inventory substrates are also available labeled with 5TAMRA. Those listed as untested were indeed previously so, but are now validated IMAP targets (during the course of this work; see text for details). The kinases range across the kinome to showcase the generic nature of the IMAP platform.


USING THE SUBSTRATE FINDER PLATES TO DETERMINE OPTIMAL PROFILER SUBSTRATE
As shown in Table 1, some of the kinases in the proposed Profiler did not have available substrates optimized for IMAP at the outset of this work. For initial assay development, usable substrates for each of these kinases will be determined with the IMAP Substrate Finder kits. The Substrate Finders enable the researcher to quickly identify suitable substrates for a target kinase. MDC offers three kits addressing different branches of the kinome as shown in Figure 1: IMAP Substrate Finder 1 for CAMK/AGC Ser/Thr Kinases (SF Ser/Thr 1), IMAP Substrate Finder 2 for CK1, TKL, STE and CMGC Ser/Thr Kinases (SF Ser/Thr 2) and IMAP Substrate Finder for Tyrosine Kinases (SF Tyr). Each kit includes 50-70 fluoresceinlabeled peptides arrayed in quadruplicate in a ready-to-use concentration on a 384-well plate (2 plates/kit). The researcher adds ATP and a single concentration of the enzyme, incubates, adds Binding S
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