Science Catalog 2012 Life Cell Signaling ADME Assays CYP450 Assay Systems Product P450-Glo™ CYP1A2 Induction/Inhibition Assay P450-Glo™ CYP3A4 Assay with Luciferin-IPA P450-Glo™ CYP3A4 Assay (Luciferin-PPXE) DMSO-Tolerant Assay P450-Glo™ CYP3A4 Assay (Luciferin-PFBE) Cell-Based/Biochemical Assay P450-Glo™ CYP1A1 Assay P450-Glo™ CYP1B1 Assay P450-Glo™ CYP1A2 Assay P450-Glo™ CYP2C8 Assay P450-Glo™ CYP2C9 Assay P450-Glo™ CYP3A4 Assay P450-Glo™ CYP3A7 Assay P450-Glo™ CYP2C19 Assay P450-Glo™ CYP2D6 Assay Available Separately NADPH Regeneration System Size Cat.# Price (Fr) 10 ml V8421 220.00 50 ml V8422 880.00 10 ml V9001 220.00 50 ml V9002 880.00 10 ml V8911 220.00 50 ml V8912 880.00 10 ml V8901 220.00 50 ml V8902 880.00 10 ml V8751 220.00 50 ml V8752 880.00 10 ml V8761 220.00 50 ml V8762 880.00 10 ml V8771 220.00 50 ml V8772 880.00 10 ml V8781 220.00 50 ml V8782 880.00 10 ml V8791 220.00 50 ml V8792 880.00 10 ml V8801 220.00 50 ml V8802 880.00 10 ml V8811 220.00 50 ml V8812 880.00 10 ml V8881 220.00 50 ml V8882 880.00 10 ml V8891 220.00 50 ml V8892 880.00 1,000 assays V9510 183.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: 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. 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 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 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 new P450-Glo™ CYP3A4 Assay with Luciferin-IPA contains a new substrate for cytochrome 3A4 that is very well suited for all applications involving human CYP3A4 and is the best substrate available for cell-based applications. Luciferin-IPA is readily taken up by cells and rapidly converted into luciferin inside the cell, which reduces the incubation time required. The low background and high signal-to-noise ratio produced using Luciferin-IPA means less starting material is required. Dimethyl sulfoxide (DMSO) can inhibit the activity of the 3A4 isoform of cytochrome P450, even at low concentrations (<0.1%).The P450-Glo™ CYP3A4 System (Luciferin-PPXE) DMSO-Tolerant Assay is designed to tolerate DMSO in the 3A4 reaction. The assay exhibits little to no change in the signal-to-background ratio in the presence of 0.2% DMSO, compared to a no-DMSO control. Features: • Obtain Reliable Results: The broad dynamic range, low background and better sensitivity result in less ambiguous data. • Avoid Fluorescence Interference: Luminescent output eliminates interference from fluorescent test compounds. • Save Time: Homogeneous assay with simple “add-and-read” format. • Avoid False Hits: Special formulation results 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/ Storage Conditions: Store the CYP1A2, CYP2C9 and CYP3A4 membranes at –70°C. Cytochrome P450 may lose activity with repeated freeze-thaw cycles. Avoid multiple freeze-thaw cycles by dispensing the CYP1A2, CYP2C9 and CYP3A4 membranes into single-use aliquots (e.g., 50μl for 96 reactions). Store aliquots at –70°C. All other components can be stored at –20°C or –70°C and protected from light. Protocol P450-Glo™ Assays Technical Bulletin P450-Glo™ substrate cytochrome P450 luciferin luciferase firefly Conversion of the P450-Glo™ substrate by cytochrome P450. Cytochrome P450 enzymes act on the P450-Glo™ luminogenic substrates to produce luciferin, a substrate for luciferase. Luciferase uses the luciferin to produce light. Light Part# TB325 20 For complete and up-to-date product information visit: www.promega.com/catalog 4125MB08_3A Pagina 23
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