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Ac-IETD-AFC is a fluorogenic substrate that can be cleaved by caspase-8 and related enzymes that recognize the amino acid sequence IETD, including caspase-3 processing enzyme, caspase-10, and granzyme B.1 Caspase activity can be quantified by fluorescent detection of free AFC (also known as 7-amino-4-trifluoromethylcoumarin), which is excited at 400 nm and emits at 505 nm.
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1. Synthesis, enzymatic evaluation, and docking studies of fluorogenic caspase 8 tetrapeptide substrates. ChemMedChem 5(1), 103-117 (2010).