| Assay Method Information | |
| | CYP450 Enzyme Induction Study |
| Description: | 500 μL of a final incubation system contains 50 μL of liver microsomes (protein concentration: 0.2 mg/mL), 1 μL of mixed CYP450 specific substrates (CYP1A2, CYP 2B6, CYP 2C9, CYP2C19, CYP 2D6, and CYP 3A4), 398 μL PBS buffer (PH7.4), 1 μL specific positive inhibitor (positive control group) or the test compound (formulated with acetonitrile), and 50 μL NADPH+MgCl2. Duplicate incubation systems of 0.5 mL each were formulated for each CYP450 subtype. A total volume of 450 μL of a uniformly mixed solution of the substrate and the enzyme was formulated in each tube, and the solution and NADPH were pre-incubated at 37° C. for 5 minutes, respectively. Then 50 μL of the mixed solution of NADPH+MgCl2 was added for reaction. 50 μL of the reaction solution was taken out at 30 minutes, and the reaction was terminated with 300 μL of ice acetonitrile containing an internal standard. In addition, two control groups of 500 μL each without NADPH were prepared in parallel as a negative control group. |
| Affinity data for this assay | |
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