货号： ADI-907-041 英文名： Boppard 数量： 1 Kit 供应商： 上海金畔生物科技有限公司-中国 规格： 瓶
The Enzo Life Sciences P450 fluroescent demethylacting activity kit is designed to quantitatively measure the enzymatic activity of formaldehyde-producing enzymes such as Cytochrome P450s. The kit is unique in that the fluorescent substrate is not involved in the multicomponent P450 reaction, but measures the product of the demethylation, formaldehyde. No separation or washing is required. The kit has been validated for 3 specific P450 systems and should work with any biological system that is producing formaldehyde as a product of demethylation.
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 (see below). 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 (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 arise from the inhibition of cytochrome P450 activities.
Lipid plays an important role in the reconstitution of P450-dependent activities after protein purification. Most in vitro studies for the reconstitution of P450 activities use dilaurylphosphatidylcholine (DLPC) as the lipid component. The reconstitution of enzymatic activity involves a concentrated incubation of P450, its redox partners (NADPH and reductase), and lipid followed by dilution into the final assay components. The reported preincubation conditions vary significantly.
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