Abcam's Caspase 8 Assay Kit (Fluorometric) provides a simple and convenient means for assaying the activity of caspases that recognize the sequence IETD. The assay is based on detection of cleavage of substrate IETD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). IETD-AFC emits blue light (?max = 400 nm); upon cleavage of the substrate by caspase 8 or related caspases, free AFC emits a yellow-green fluorescence (?max = 505 nm), which can be quantified using a fluorometer or a fluorescence microtiter plate reader. Visit our FAQs page for tips and troubleshooting.
Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells.
Caspases are cysteine proteases, expressed as inactive precursors, that mediate apoptosis by proteolysis of specific substrates. Caspases have the ability to cleave after aspartic acid residues. There are two classes of caspases involved in apoptosis; initiators (activation by receptor cluster) and effectors (activation by mitochondrial permeability transition). Proapoptotic signals autocatalytically activate initiator caspases, such as Caspase 8 and Caspase 9. Activated initiator caspases then process effector caspases, such as Caspase 3 and Caspase 7, which in turn cause cell collapse.