Caspase-5 Fluorometric Assay Kit

Caspase-5 belongs to the Caspase-family of cysteine proteases. Caspase-5 is an enzyme that proteolytically cleaves other proteins at an aspartic acid residue. Caspase-5 exists in cells as an inactive proenzyme and is matured by proteolysis. The active Caspase-5 is a heterotetramer containing two large and two small subunits. The function of Caspase-5 is believed to be an inflammatory Caspase, together with Caspase-1, Caspase-4, and has a role in the immune system.

Apoptosis in mammalian cells is caused by activation of ICE-family proteases/caspases. The Caspase-5 Fluorometric Assay Kit provides a fast and convenient means to assay the activity of Caspase-5 and other related caspases. These caspases can recognize the sequence WEHD. The assay is according to the detection of cleavage of substrate WEHD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). WEHD-AFC emits blue light (λmax = 400 nm); free AFC emits a yellow-green fluorescence ( λmax = 505 nm) upon cleavage of the substrate by Caspase-5 or other related caspases. Using a fluorometer or fluorecence microtiter plate reader can quantify the fluorescence. Comparison of the fluorescence of AFC from a treated sample with an untreated control allows determination of the fold increase in Caspase-5 activity.