| Step | Parameter | Value | Description |
|---|---|---|---|
| 1 | Compound | 0.5 µL | DMSO 50%, HTRF® Enzymatic buffer 1x |
| 2 | Enzyme | 5.5 µL | Dilution in HTRF® Enzymatic buffer 1x |
| 3 | Incubation | 15min | Room temperature, covered with plate seal |
| 4 | Substrate | 2 µL | Dilution in HTRF® Enzymatic buffer 1x |
| 5 | ATP | 2 µL | Dilution in HTRF® Enzymatic buffer 1x |
| 6 | Incubation | 10 or 30min | Room temperature, covered with plate seal |
| 7 | Reagent | 10 µL | Europium Anti-P-MAb, SA-XL665 in HTRF® Detection buffer |
| 8 | Incubation | 60 min | Room temperature |
| 9 | Detection | HTRF® mode | Rubystar plate reader |