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. 2017 Aug 31;39(2):302–310. doi: 10.1038/aps.2017.59

Figure 4.

Figure 4

FP competitive binding assay and determination of the Z' factors. (A) Selection of the EED concentration for the FP competitive binding assay based on the FP saturation binding curve at 80% fractions of tracer bound. (B) FP competitive binding curve. Mixtures of 20 nmol/L FITC-EZH2(40-63) and 625 nmol/L EED were incubated in the presence of different concentrations of unlabeled peptide EZH2(40-63) (20 nmol/L to 40 μmol/L). The FP values were recorded at 2 h. (C–F) Z' factors for evaluation of the FP-based high-throughput screening assay in the absence or presence of 1%, 2%, and 4% DMSO. The FP values of both negative and positive controls are shown in a scatter plot. All experiments were performed at least three times. The data were analyzed in GraphPad Prism 5.0.