FP vs. TR-FRET Detection

Akt inhibition with staurosporine: Comparison of IMAP TR-FRET (top) and IMAP FP (bottom) detection. Reaction conditions: Akt kinase (0.1u/ml, Upstate: 14-276), 300 nM, 1µM, 3µM FAM-Crosstide (5FAM-GRPRTSSFAEG-COOH, R7110), 10 µM ATP, Staurosporine as indicated. IMAP Binding Solution: 40% Binding Buffer A, 60% Binding Buffer B, Progressive Binding Reagent 1:400.