Ac-IETD-AFC is a fluorogenic substrate of caspase-8, caspase-10 and granzyme B. Working concentration of this substrate is 25-50 µM. The released AFC (7-Amino-4-trifluoromethylcoumarin) fluorescence can be detected by a fluorimeter or a plate reader using excitation/emission wavelengths at 400 nm/505 nm, respectively.