The CasPASE™ Apoptosis assays are designed to monitor apoptosis by measuring various caspase (protease) activities, a key early indicator of apoptosis in mammalian cells. The assay provides a simple and easy to follow method that can be monitored with a fluorescence reader.
The assay is based on the detection of cleavage of a synthetic substrate, which has 7-amino-4-trifluromethyl coumarin (AFC) at the C-terminal. When liberated from the peptide, AFC produces an optical change that exhibits a fluorescence spectral shift between the substrate-conjugate and the free dye. The AFC substrate is fluorogenic (detected at 480-520 nm with a fluorometer). During the reaction, the substrate releases AFC free dye and undergoes a fluorescence shift.
Comparison of the fluorescence from an induced/apoptotic sample with an uninduced control allows one to determine the fold-increase in protease activity. When measuring fluorescence, the assay reaction is excited at 380 to 400nm and emission is read at 480 to 520nm.
The assays 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.
Colorimetric Caspase Assays are also available.
Features
- Assays for multiple caspase enzymes are available
- Exhibit fluorescence spectral shift
- Conveniently adaptable for high-throughput 96-well format
Applications
- Monitor apoptosis by measuring various caspase activities in cells and tissues
- Use with purified enzyme preparations, cell extracts, and tissue lysates
- Characterization of caspase enzymes