General Description
Caspase-13 Assay Kit, Fluorometric with Ac-LEED-AFC substrate
Caspase assay kits are designed to assess various protease/caspase activities as an early indicator of apoptosis initiation in mammalian cells. These assay kits provide a simple and convenient way to monitor caspase activities with a fluorescence reader. Assay principle is based on the detection of cleavage of a synthetic amino acid sequence bearing 7-amino-4-trifluromethyl coumarin (AFC) at the C-terminal end. Following cleavage from the peptide, the substrate releases AFC free dye and undergoes a fluorescence shift to generate an optical change with a fluorescence spectral shift between the substrate-conjugate and the free dye, detectable at 510 to 550 nm with a fluorometer.
For fluorescence measurement, the excitation is at 380 to 400 nm and emission is measured at 510 to 550 nm. The assays can be conveniently adapted for high-throughput multi-well plate format, and is compatible with purified enzyme preparations, cell extracts or tissue lysates.
