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. 2010 Feb;8(1):47–62. doi: 10.1089/adt.2009.0212

Fig. 6.

Fig. 6.

Plk1 time-resolved fluorescence energy transfer (TR-FRET) assay with T210D mutant. (A) Enzyme titration in a 384-well plate format. Indicated concentration of Plk1ΔC (◯) and Plk1T210D (●) was incubated with ATP and b-FKD for 60 min at 25°C. The assay plate received EDTA/TR-FRET, and was loaded onto the Victor2™ after 45 min. Error bars demonstrate standard error of the medians for n = 15. (B) Enzyme titration in the assay in 1,536-well plate. Indicated concentration of Plk1T210D was incubated in 60 (◯) and 120 (●) min. Error bars demonstrate standard error of the medians for n = 44. (C) Dose–response to the known inhibitors. Indicated concentration of staurosporine (left) and K252a (right) was added to the 60-min reaction with 4 nM Plk1ΔC (◯) and 66 nM Plk1T210D (●). Error bars demonstrate standard error of the medians for n = 8.