Specifications

  • Catalog Number K011-F
  • Assay Type Activity Assay
  • Sample Types Any P450 system or biological system producing demethylation product, Formaldehyde
  • Assay Duration 45 Minutes
  • Samples/Plate 41 in Duplicate
  • Readout Fluorescent, 510 nm Emission / 450 nm Excitation

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Assay Principle:

The DetectX® P450 Demethylation Fluorescent Activity Kit provides a reliable method for quantitatively measuring demethylating P450 system activity. Suitable for any P450 system or biological system producing Formaldehyde as a demethylation product, this kit features a 45 to 90 minute run time. Detailed instructions are available in the kit insert. Please read instructions thoroughly before starting the assay. The kit includes a Formaldehyde Standard for generating a precise standard curve.

Protocol Summary:

  • Add standards or diluted samples to the black microtiter plate.
  • Introduce NADPH and incubate at 37°C for 15-60 minutes to facilitate the P450 demethylase reaction.
  • Add DetectX® Formaldehyde Detection Reagent to each well, ensuring proper mixing.
  • Incubate the mixture at 37°C for 30 minutes, allowing the fluorescent reaction between the Detection Reagent and the Formaldehyde produced in the P450 demethylase reaction.
  • Use a plate reader to detect the fluorescent signal at 510nm and calculate P450 demethylase activity using the standard curve.

Background:

Cytochrome P450 enzymes, present in most tissues, play important roles in various biological functions such as hormone synthesis and breakdown, cholesterol synthesis, and vitamin D metabolism. They are also key players in drug metabolism, influencing the development of new pharmaceuticals and drug interactions. As a superfamily of heme-containing enzymes, cytochrome P450s exhibit significant diversity in substrate specificity and catalytic activity.

These enzymes metabolize a wide range of exogenous and endogenous compounds, participating in multicomponent electron transfer reactions. In toxicology, cytochrome P450 enzymes are of particular interest as they break down toxins, preventing harmful accumulation in the body. Their involvement extends to the metabolism of environmental chemicals and carcinogens and determines the sensitivity of individuals to various environmental toxins and drugs.

The DetectX® P450 Demethylation Fluorescent Activity Kit provides a sensitive and straightforward approach to assessing the activity of these enzymes, facilitating research in drug development, toxicology, and understanding the enzymatic mechanisms underpinning critical physiological processes.

P450 Demethylation Fluorescent Activity Kit

$386.00

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The DetectX® P450 Fluorescent Activity Kit allows activity measurement of any demethylating P450 system WITHOUT additions to the P450:Substrate reaction by measuring the formaldehyde generated by demethylation.

SKU: K011-F1 Category: