Figure 3. Assay robustness for chemical screening.
(A) Repeated (n=3) 384-well plate high- to-low controls were performed for SET7/9, SET8, SETD2, and EuHTMase1. For the high controls, the wells contained 2 μl of 9:1 H2O/DMSO prior to the addition of PMTs, substrate and cofactor. In comparison, the wells of the low controls contained 2 μl of 100 mM HClO4. Lowering pH inactivated PMTs. (B) Assay for the four enzymes. SET7/9, SET8, SETD2 and EuHMTase1 were evaluated on percent coefficient of variation (%CV) and Z′ factor.