The CasPASE^™ Apoptosis assays are designed to monitor apoptosis by measuring caspase activity, an early indicator of apoptosis. The assay can be monitored with a fluorescence reader, spectrophotometer, or a titer plate reader.

 

The kit uses AFC-substrate (7-amino-4-trifluromethyl coumarin conjugated at C-terminal), which exhibits both fluorescence and absorbance spectral shifts between the substrate-conjugate and the free dye (AFC). The AFC substrate is both chromogenic (yellow-green color visible to the naked eyes) and fluorogenic (detected at 510-550nm with a fluorometer). During the reaction, the substrate releases AFC free dye and undergoes both a fluorescence and absorbance shift. Since AFC-substrate exhibits dual-mode detection capability, arbitrary fluorescence values can be standardized to an optical density value.

 

When measuring fluorescence, the assay reaction is excited at 380-400nm and emission is read at 510-550nm. If the assay is measured with a spectrophotometer or a titer plate reader, the absorbance is measured at 380-400nm.

 

The assay can be conveniently adapted for high-throughput 96-well format. The assay system may be used with purified enzyme preparations, cell extracts or tissue lysates.

 

CasPASE^™ assays are available for caspase enzymes 1-10 and 13. Each CasPASE^™ assay kit is supplied with necessary assay buffers, enzyme specific AFC-substrate, free dye (AFC) and the potent caspase inhibitor Z-VAD-FMK for establishing proper positive and negative controls and standards. Available in a 50 or 100 assay size.