| Assay Method Information | |
| | CYP450 Enzyme Induction Study |
| Description: | A final incubation system of 500 μL contained 50 μL of liver microsomes (protein concentration: 0.2 mg/mL, Corning), 1 μL of mixed specific substrates of CYP450 (CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP3A4), 398 μL of PBS (pH 7.4), 1 μL of specific positive inhibitor (positive control) or test compound (in acetonitrile) and 50 μL of NADPH+MgCl2. Samples were prepared in duplicate of 0.5 mL for each CYP450 subtype. For each tube, the 450 μL mixed solution of substrates and enzyme and the NADPH solution were separately pre-incubated at 37° C. for 5 min. The 50 μL mixed solution of NADPH+MgCl2 was added for reaction. At 30 min, 50 μL of the mixture was taken and 300 μL of glacial acetonitrile containing an internal standard was added to terminate the reaction. Additionally, 2 blanks of 500 μL each were prepared in parallel without adding NADPH as the negative control group. |
| Affinity data for this assay | |
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