Culture medium | Williams’ E medium without phenol red, containing 2 mM glutamine and 15 mM Hepes |
Incubation | 37°C in 5% CO2/95% air, humidified atmosphere under agitation (450 rpm) |
Hepatocytes suspension prewarming | 30 min at 2 million/ml in similar conditions as incubation |
Final incubation volume | 500 µl |
Final hepatocyte concentration | 1 million/ml |
Substrate concentration | 0.5 µM |
Azamulin concentration | 3 µM |
Final organic solvent concentration | 1% ACN |
Incubation time | 4 h |
Sampling time | 0, 30, 60, 120, 180, and 240 min |
Sampling volume | 50 µl/time point |