| Assay Method Information | |
| | CYP Enzyme Single Point Inhibition Assay |
| Description: | 1. Experimental ObjectiveThe human liver microsome incubation system was used to rapidly predict the inhibitory effect of the compounds on CYP450 enzyme subtypes by using single-point method.2. Experimental Process2.1 Formulation of SolutionNADPH (reduced nicotinamide adenine dinucleotide phosphate) was weighed, to which 100 mM phosphate buffer was added to obtain a final concentration of 2.5 mM. 4 mL of 100 mM phosphate buffer was added to 50 μL of 20 mg/mL microsomes, and mixed well to obtain 0.25 mg/mL microsomes.Formulation of test compound reaction solution:The compound of the example to be tested was weighed, diluted to 10 mM with DMSO, and then diluted to 100 μM with 100 mM phosphate buffer.2.2 Experimental Procedure1. 40 μL of liver microsomes, 10 μL of substrate, and 10 μL of the test compound were added to a 96-well plate, and pre-incubated for 3 minutes.2. 40 μL of NADPH was added.3. 300 μL of acetonitrile stop solution containing an internal standard was added at 20 minutes.4. The sample was injected by centrifuge. |
| Affinity data for this assay | |
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