Product category:
Assay kits
News Release from: Promega UK | Subject: P450-Glo CYP450 assays
Edited by the Laboratorytalk Editorial
Team on 20 October 2003
Luminescent assays for sensitivity and
specificity
A simple and fast means of measuring the effects of analytes - such as potential new drugs - on the activity of human cytochrome P450s
By employing luminogenic P450 substrates derived from beetle luciferin, the new P450-Glo CYP450 assays from Promega provide a simple and fast means of measuring the effects of analytes - such as potential new drugs - on the activity of human cytochrome P450s The homogenous luminescent method used in these assays result in each having a low background level and a large dynamic range
This article was originally published on Laboratorytalk on 4 Sep 2002 at 8.00am (UK)
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Sensitivity is exceptional, allowing the use of less cytochrome P450 compared with other HPLC or fluorometric methods and hence reducing overall assay cost.
All of the assays are suitable for investigating activity of P450s from both recombinant and native sources.
A stable signal with a half-life of more than two hours allows batch plate processing and the assays are easily scalable to 384-well plate format (Z' values are greater than 0.8 in either 96- or 384- well plate formats).
Unequivocal results can be obtained because there is no fluorescent excitation or emission overlaps from any of the assay components or analytes.
In addition, the stabilised firefly luciferase and a proprietary buffer formulation minimises false positives when screening for cytochrome P450 inhibitors.
The luciferin derivatives used in the assays cannot be metabolised by luciferase.
Instead, they are converted to luciferin (the luciferase substrate) by cytochrome P450 so that the luminescence produced is directly proportional to the level of P450 activity.
The speed and simplicity of P450-Glo CYP450 assays add-and-read format makes them ideal for screening drugs and new chemical entities for any P450 effects on activity very early in their development.
This should therefore reduce the likelihood of incurring the delay and expense of late stage failures.
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