| Assay Method Information | |
| | Test on EZH2 (wt) Inhibitory Activity |
| Description: | Table 4: ProceduresCompounds at different concentrations were dissolved in 100% DMSO and transferred to test plates, and the final concentration of DMSO was 1%. Enzyme solution and substrate SAM solution were prepared with 1-fold reaction buffer (optimized Tris buffer). 5 uL of enzyme solution was added to the test plate and wells only added with 5 uL of reaction buffer were used as negative control wells. After incubation at room temperature for 15 min, the plate was added with 5 uL of substrate solution per well to initiate the reaction, and then incubated at room temperature for 60 min. 5 uL of receptor solution was added and the plate was incubated away from light at room temperature for 60 min. 10 uL of donor solution was added and the plate was incubated away from light at room temperature for 30 min. The endpoint values were read on the Envision and the IC50 was calculated.Data CalculationData were fitted by GraphPad Prism 5 software and inhibition rate was calculated according to equation (1)Inh %=(Max−Signal)/(Max−Min)×100 (1)IC50 was calculated using equation (2)Y=Bottom+(Top−Bottom)/(1+10{circumflex over ( )}((Log IC 50 −X)×Hill Slope))Y: inhibition rate; X: concentration of the compound |
| Affinity data for this assay | |
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