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Assay Method Information |
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| | Biochemical Assay |
| Description: | Each compound was dissolved in DMSO to a concentration of 10 mM and further diluted to 100 μM using acetonitrile. Liver microsomes from selected species were incubated in duplicate with each compound at a final concentration of 1 μM in 0.1 M potassium phosphate buffer (pH 7.4) containing 3.3 mM MgCl2, 0.5 mg/ml microsomal protein, in the presence or absence of NADPH (1 mM). Incubations were performed at 37° C. Control incubations with reference substances were included for each experiment. At different time points (t=0, 5, 15, 30, 45 min), an appropriate aliquot of the incubation mixture was transferred into a quench plate containing acetonitrile and internal standard cooled to 4° C. After the last time point, the quench plates were mixed thoroughly and centrifuged for 15 minutes at 3700 rpm and 10° C. (Eppendorf 5804R). The supernatant was transferred to new 96 well plates and subjected to LCMS analysis. The disappearance of the parent compound was determined. |
| Affinity data for this assay | |
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