Ready-to-use fluorometric substrate for caspase-4 and related caspases that recognize the amino acid sequence LEVD. Caspase-4 and related caspase activity can be quantified by fluorescent detection of free AFC after cleavage from the peptide substrate LEVD-AFC at Ex. = 400 nm and Em. = 505 nm, using a fluorometer or multi-well fluorescence plate reader. Alternatively, a shift in fluorescence from blue to green upon cleavage can be visualized using a hand-held long-UV lamp.