Table 1.
Inhibitor IC50 Values for Direct and Cascade Kinase Assays. Kinase Reactions were Conducted in the Presence of Increasing Inhibitor Concentrations for the FRET Direct, FRET Cascades, and TR-FRET Direct Assays Using 100 μM ATP
| Inhibitor IC50 (nM) | ||||||
|---|---|---|---|---|---|---|
| Format | Assay | Staur. | GW5074 | ZM336372 | U0126 | PD98059 |
| FRET Direct | ERK2 | 4,400 | >400 | >50,000 | >50,000 | >50,000 |
| FRET Double | MEK1-ERK2 | 4 | >400 | >50,000 | >10,000 | >50,000 |
| FRET Triple | BRAF-MEK1-ERK2 | 56 | 3 | 400 | 65 | 420 |
| FRET Triple | BRAF V599E-MEK1-ERK2 | 69 | 2 | 32 | 65 | 300 |
| FRET Triple | CRAF-MEK1-ERK2 | 63 | 9 | 30 | 66 | 380 |
| TR-FRET Direct | BRAF | >50,000 | 15 | 580 | >40,000 | >50,000 |
| TR-FRET Direct | BRAF V599E | >50,000 | 4.9 | 270 | ≥5,000 | >50,000 |
| TR-FRET Direct | CRAF | >50,000 | 24 | 90 | >50,000 | >50,000 |