Efficiently detect active caspase-3 in mammalian cells undergoing apoptosis by using the Caspase-3/7 Activity Flow Cytometry Kit, Green. This reagent kit uses a quick and simple staining protocol that is highly sensitive and compatible with standard flow cytometers.

Caspase-3 and caspase-7 have a proteolytic substrate selectivity for the peptide sequence Asp-Glu-Val-Asp (DEVD). This kit includes TF2-DEVD-FMK—a cell-permeable and non-toxic fluorescent substrate—as well as an assay buffer and propidium iodide. When added to apoptotic cells, TF2-DEVD-FMK irreversibly binds to activated caspase-3, resulting in the retention of the green fluorescent reagent within cells that can be detected by flow cytometry.

Caspases are a family of proteases that play a central role in cellular apoptosis through the cleavage of select proteins, and result in the disassembly of the cell (Thornberry & Lazebnik Science, 1998). This process is switched on by pro-apoptotic signals from death receptors or stimuli such as cytotoxic reagents, which activate the initiator caspases and lead to activation of the effector caspases—caspase-3 and caspase-7 (Thornberry & Lazebnik Science, 1998). Caspase-3/7 Activity Flow Cytometry Kit, Green is useful for screening caspase-3 inhibitors and quantifying apoptotic cells containing active caspase-3.