CYP1A2 Assay Systems

The P450-Glo™ CYP450 Assays provide a homogeneous, luminescent method for measuring cytochrome P450 activity. The assays are designed to measure the activities of P450s from recombinant and native sources and for testing the effects of analytes such as drugs and new chemical entities on P450 activities. These luminescent assays exhibit exquisite sensitivity, low background signals and broad dynamic range.

P450-Glo™ Assays employ luminogenic P450 substrates that are derivatives of beetle luciferin, a substrate for luciferase enzymes. The derivatives are not substrates for luciferase but are converted by P450s to...

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The P450-Glo™ CYP450 Assays provide a homogeneous, luminescent method for measuring cytochrome P450 activity. The assays are designed to measure the activities of P450s from recombinant and native sources and for testing the effects of analytes such as drugs and new chemical entities on P450 activities. These luminescent assays exhibit exquisite sensitivity, low background signals and broad dynamic range.

P450-Glo™ Assays employ luminogenic P450 substrates that are derivatives of beetle luciferin, a substrate for luciferase enzymes. The derivatives are not substrates for luciferase but are converted by P450s to luciferin, which in turn reacts with luciferase to produce light that is directly proportional to the activity of the P450.

The P450-Glo™ Assays generate a "glow-type" luminescent signal, produced using derivatized luciferins as P450 substrates and a recombinant stabilized luciferase (Ultra-Glo™ Luciferase) coupled with a proprietary buffer system. The half-life of the luminescent output is greater than two hours, eliminating the need for luminometers with injectors and allowing for batch plate processing. The formulation also minimizes the incidence of false positives due to inhibition of luciferase by analytes when screening for cytochrome P450 inhibitors.

The P450-Glo™ CYP1A2 Induction/Inhibition Assay contains a new substrate for cytochrome 1A2 that is very well suited for all applications involving human CYP1A2 and is the best substrate available for cell-based applications. The substrate, Luciferin-1A2, is readily taken up by cells and converted into a luciferin precursor by CYP1A2. The luciferin precursor is rapidly converted into luciferin following the addition of D-cysteine. After addition of luciferin-1A2 to cells, the assay can be completed in approximately one hour. The low background and high signal-to-noise ratio produced using Luciferin-1A2 means less starting material is required.

The P450-Glo™ CYP1A2 Screening System provides a complete set of reagents for performing luminescent CYP1A2 assays using recombinantly expressed CYP1A2. The system includes a membrane preparation containing recombinant human cytochrome 1A2, a luminogenic cytochrome P450 substrate (luciferin-ME), an NADPH Regeneration System, reaction buffer, Luciferin Detection Reagent and Luciferin-Free Water. The membranes are prepared from baculovirus-infected insect cells and contain human cytochrome P450 and P450 reductase. The P450-Glo™ Screening System also contains a membrane fraction devoid of cytochrome P450 activity as a negative control. The assay is ideal for testing the effects of drugs and new chemical entities on cytochrome P450 enzyme activities.

Features - Benefits

• Complete Systems: The screening system includes a membrane preparation containing recombinant human CYP1A2 enzyme, a luminogenic cytochrome P450 substrate (luciferin-ME), an NADPH regeneration system, reaction buffer, Luciferin Detection Reagent and Luciferin-Free Water.
• Excellent Selectivity:The P450-Glo™ CYP1A2 Induction/Inhibition Assay contains Luciferin-1A2, a pro-luciferin substrate that is more selective for CYP1A2, which makes it ideal for use in cell-based applications.
• Obtain Reliable Results: The broad dynamic range, low background and better sensitivity result in less ambiguous data.
• Save Time: Homogeneous assay with simple "add-and-read" format.
• Avoid False Hits: Special formulation and luminescent signal result in low false-hit rate.
• Save Money: Scalable to 384-well format, reducing cost per well.
• Automate This Assay: Validated automated methods available at: www.promega.com/automethods/
• Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/

Applications

• Measure recombinant CYP1A2 activities in membrane fractions from heterologous expression systems, such as Sf9 cells and E. coli.
• Measure native CYP1A2 activity in cells and microsomal fractions from cells and tissues.
• Screen drugs and new chemical entities for their capacity to modulate CYP450 activities in native or recombinant fractions.

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