Employs the fluorescent inhibitor probe SR-LEHD-FMK to label active caspase-9 enzyme in living cells or tissue samples.
Caspases play important roles in apoptosis and inflammation. FLICA® assay kits are used by researchers seeking to quantitate apoptosis via caspase activity in cultured cells and tissues. The FLICA reagent SR-LEHD-FMK enters each cell and irreversibly binds to activated caspase-9. Because the SR-LEHD-FMK FLICA reagent becomes covalently coupled to the active enzyme, it is retained within the cell, while any unbound SR-LEHD-FMK FLICA reagent diffuses out of the cell and is washed away. The remaining red fluorescent signal is a direct measure of the active caspase-9 enzyme activity present in the cell at the time the reagent was added. Cells that contain the bound FLICA can be analyzed by a fluorescent plate reader, fluorescence microscopy, or flow cytometry. Cells labeled with the FLICA reagent may be read immediately or preserved for 24 hours using the fixative. Unfixed samples may also be analyzed with Hoechst stain to detect changes in nuclear morphology.
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