Caspases play important roles in apoptosis and cell signaling. The activation of caspase-3 (CPP32/apopain) is important for the initiation of apoptosis. Caspase 3 is also identified as a drug-screening target. Caspase 3 has substrate selectivity for the peptide sequence Asp-Glu-Val-Asp (DEVD). This Amplite™ Caspase-3 Assay Kit uses (Z-DEVD)2R110 as fluorogenic indicator for assaying caspase-3 activity. Cleavage of R110 peptides by caspases generates strongly fluorescent R110 that can be monitored fluorimetrically at 510-530 nm with excitation of 488 nm, the most common excitation light source used in fluorescence instruments. R110-derived caspase substrates are probably the most sensitive indicators widely used for the fluorimetric detection of various caspase activities. This kit can be used to continuously measure the activities of caspase-3 in cell extracts and purified enzyme preparations using a fluorescence microplate reader or fluorometer. It can also be used with flow cytometry for analyzing cell apoptosis and the activities of caspases 3 and 7.
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