The Caspase-Glo® 3/7 assay provides a proluminescent caspase-3/7 DEVD-aminoluciferin substrate and a proprietary thermostable luciferase in a reagent optimized for caspase-3/7 activity, luciferase activity and cell lysis. Adding the single Caspase-Glo® 3/7 Reagent in an "add-mix-measure" format results in cell lysis, followed by caspase cleavage of the substrate. This liberates free aminoluciferin, which is consumed by the luciferase, generating a "glow-type" luminescent signal that is proportional to caspase-3/7 activity.
The stabilized luciferase and proprietary buffer system improve assay performance across a wide range of assay conditions, and the assay is less likely to be affected by compound interference than fluorescent- or colorimetric-based assays. The caspase activity assay is designed for use with multiwell plate formats using either purified enzyme or cells in culture and can be multiplexed with other cell-based assays. The assay delivers excellent Z´-factor values in cell and purified enzyme models.