James J. Cali, Mary Sobol, Dongping Ma, Susan Frackman,
Promega Corporation, 2800 Woods Hollow Rd., Madison, WI 53711
P450-Glo™ Luminescent P450 Assays utilize derivatives of beetle luciferin as P450 enzyme substrates. Luciferin is a substrate for the light generating reaction of firefly luciferase. The derivatives are inactive with luciferase but are metabolized by P450s to luciferin, which in turn reacts with luciferase to make light. A homogenous luciferase mixture is added directly to P450 reactions that use luciferin derivatives as substrates. The amount of luciferin produced by a P450 is directly proportional to light output so light is used to measure P450 activity. Substrates are provided for CYP1A1, 1A2, 1B1, 2C8, 2C9, 2C19, 2D6, 2J2, 3A4, 3A7, 4A11, 4F3B, 4F12 and 19. P450-Glo™ Assays detect P450 activities in recombinant preparations, liver microsomes and cultured cells. P450-Glo™ Assays provide a simple, homogenous and robust luminescent assay format for rapid screening of multiple compounds.