Fig. (5) Direct TR-FRET MAP3K assays. (A) The TR-FRET ratios (Em 520 nm/Em 495 nm) resulting from titrations B-RAF V599E with one of five different ATP concentrations (1 – 1000 µM). (B) The TR-FRET direct assay was utilized with seven different MAP3Ks using a final ATP concentration of 500 µM. All data points represent the mean ± standard deviation from triplicate samples.