Mouse P450 Fluorescent Activity kit allows activity measurement of any demethylating P450 system WITHOUT any additions to the the P450:Substrate reaction. This assay measures the formaldehyde generated by demethylation and the signal is read AFTER the P450 reaction has been terminated. Convenient plate assay with 30 minute fluorescent substrate incubation and detection readout at 510 nm. Tested in 3A4, 2D6 and 2B4 P450 systems with erythromycin, dextromethorphan or benzphetamine.
The cytochrome P450s (P450s) are a superfamily of heme containing enzymes that display tremendous diversity with regard to substrate specificity and catalytic activity. P450s use a plethora of both exogenous and endogenous compounds as substrates in their reactions. Usually they form part of multicomponent electron transfer reactions. Catalysis by the eukaryotic P450 enzymes involves a multistep reaction cycle that includes two steps in which electron transfer is accomplished from a redox partner. The diflavin protein, NADPH cytochrome P450 reductase contains both FAD and FMN and can transfer both electrons needed for the catalytic cycle. In some P450 reactions, the second electron of the reaction cycle also can be delivered by cytochrome b5. The P450 enzymes and cofactors of the mammalian drug-metabolizing system are embedded in the membrane of the endoplasmic reticulum. The P450s play a crucial role in the development of new drug entities as drug interactions commonly inhibit cytochrome P450 activities.
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