| Assay Method Information | |
| | Measurement of CYP2E1 Enzymatic Activity |
| Description: | CYP2E1 hydroxylation of p-nitrophenol was determined by the spectrophotometric measurement of 4-nitrocatechol. Briefly, reaction mixture consisting of 100 mM phosphate buffer (pH 7.4), 100 uM p-nitrophenol, 2 mg/mL CYP2E1, and inhibitors were pre-incubated for 10 min at 37 deg C. Reactions were initiated by adding NADPH at a final concentration of 1 mM and were incubated for an additional 30 min at 37 deg C. Reactions were stopped upon the addition of perchloric acid (0.6 N or 22%). The effects of the HO inhibitors on CYP2E1 activity were tested at concentrations in the range of 0.01-100 uM. |
| Affinity data for this assay | |
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