Newsletter 6/2009

Luminos Interactive Assay Solutions P450

DetectX^™P450 Activity Kit

Catalog Number K011-F1

• Easy, sensitive fluorescent P450 activity
• Direct correlation to Nash reaction using less P450
• Run your P450 reaction - stop - measure HCHO generated
• Stable reagents run at one temperature

The DetectX^™ P450 Fluorescent Activity kit allows activity measurement of any demethylating P450 system using less than half the amount of P450 enzymes the Nash reaction uses and without changing the P450:Substrate:Drug interaction. This assay measures the formaldehyde generated by the demethylation and the signal is read AFTER the P450 reaction has been terminated. Convenient plate assay with fluorescent detection readout at 510 nm.

The cytochromes P450 (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 enzymatic 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-drug interactions commonly inhibit cytochrome P450 activities.

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