Wash Method | Add Method | Dilution Method | |
---|---|---|---|
Preincubation | |||
Volume (μl) | 250 | 100 | 50 |
Cell density (cells/ml) | 0.3 × 106 | 0.3 × 106 | 0.6 × 106 |
Inhibitor concentrationa (μM) | 0.1–300 | 0.1–300 | 0.1–300 |
Preincubation timeb (min) | 0–60 | 0–60 | 0–60 |
Incubation | |||
Volume (μl) | 220 | 110 | 250 |
Cell density | 0.245 × 106 | 0.245 × 106 | 0.125 × 106 |
Inhibitor concentration (μM) | 18-Fold dilution | 1.25-Fold dilution | 5-Fold dilution |
Midazolam concentration (μM) | 30 μM | 30 μM | 30 μM |
Incubation time (min) | 10 | 10 | 10 |
↵a,b Concentration and preincubation time varied depending on the inhibitor and experimental design.