The Cytochrome P450 Activity kit is designed to measure the enzymatic activity of demethylation processes carried out by Cytochrome P450. The unique feature of the Kit is that the fluorescent substrate is not involved in the multi component P450 reaction, but measures the product of the demethylation, formaldehyde. The kit has been validated for a specific P450 system and should work with any P450 system that via demethylation is producing formaldehyde as a product. The formaldehyde substrate is added after the P450 reaction has been terminated, and the measured formaldehyde is then read at 510 nm with excitation at 450 nm.
The kit provides an optimized buffer for 2B4 P450, NAPDH, a stable formaldehyde standard, the Formaldehyde Detection Reagent and a suitable 96 well plate for detecting the generated fluorescent signal. The end user will have to provide the P450 enzyme system or microsomal preparations necessary for activity, along with any inhibitors or activators. The end user should carry out the P450 reaction in our supplied reaction buffer or similar using optimized conditions for the reaction.
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