Description
Efficiently detect caspase-3/7 activity in cell lysates prepared from mammalian cells undergoing apoptosis by using the Caspase-3/7 Activity Plate Reader Assay Kit, Blue, which includes the fluorogenic substrate Ac-DEVD-AMC and an assay buffer. This reagent kit uses a quick and simple staining protocol that is well suited for high-throughput screening and compatible with standard fluorescent plate readers.
Caspase-3 and caspase-7 have a proteolytic substrate selectivity for the peptide sequence Asp-Glu-Val-Asp (DEVD). When added to apoptotic cells, Ac-DEVD-AMC is cleaved by activated caspase-3/7, generating a blue fluorescent signal that can be detected by standard plate readers.
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 assays are useful for quantifying caspase-3/7 activity in apoptotic cells and for screening caspase-3/7 inhibitors.
