Homogeneous Luminescent Caspase Assays to Detect Apoptosis and in Vitro Toxicity...
Homogeneous Luminescent Caspase Assays to Detect Apoptosis and in Vitro Toxicity Scientific Poster
Terry Riss, Martha O’Brien, Andrew Niles, Rich Moravec, Marni Amburn, Deborah Bishop, Mary Sobol, Kay Rashka, Bill Daily* and Mike Scurria*
Promega Corporation, 2800 Woods Hollow Road, Madison WI 53711 and *Promega Biosciences, 277 Granada Drive, San Luis Obispo, CA 93401
Luminogenic substrates have been developed for the measurement of caspase-3/7, -8, & -9 activities as in vitro markers of apoptosis as well as for screening for caspase inhibitors. The homogeneous format of the Caspase-Glo™ Assays was enabled by utilizing a mutant form of beetle luciferase that is stable in the conditions necessary to lyse cells and maintain caspase activity for hours. The Caspase-Glo™ Assay procedure is to add reagent directly to cells in multiwell plates, incubate for 30 min-1 h and record luminescence. There is a linear relationship between the luminescent signal and caspase activity. The luminescent format is 50-fold more sensitive than fluorescent assays for detection of recombinant caspase-3 and can detect as few as 20 apoptotic cells. The stability of the reagents and glow-type luminescent signal are compatible with automation for HTS. Multiplexing of luminescent and fluorescent caspase assays is possible.
- Part# PS015
- Printed in USA.