| Assay Method Information | |
| | P450 Inhibition Assay |
| Description: | A range of concentrations of each compound in a 96-well plate were preincubated in buffer containing recombinant human CYP450 microsomal protein and the substrate. Following preincubation, NADPH generating system (glucose-6-phosphate, NADP, and glucose 6-phosphate dehydrogenase) was added to each well to start the reaction. Production of fluorescent metabolite was measured using Cytofluor (Perkin-Elmer, US) plate reader. The rate of metabolite production (AFU/min) was determined for each concentration of compound and converted to a percentage of the mean control rates using Cytofluor software. The inhibitory potential (IC50) of each compound was determined from the slope of the plot using GraFit v5.08 (Erithacus software, UK). Miconazole was included as a positive control for each isozyme. |
| Affinity data for this assay | |
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