Calpain Activity Fluorometric Assay Kit

Apoptosis is one of the biological processes that activation of calpain is implicated in. Cell membrane and Ca²⁺ is required for calpainactivation. The activated calpain is entered into cytosol. The Calpain Activity Assay Kit is a convenient way of measuring calpain activity with optimized buffers and reagents. The Extraction Buffer extracts cytosolic proteins with contaminations from cell membrane and lysosome proteases. It also protects the cell from atuo-activation of calpain during extraction process. Thus, the kit only detects activated calpain in cytosol following treatment of cells with chemicals or drugs inducers. The fluorometric assay is based on the detection of cleavage of calpain substrate Ac-LLY-AFC. Ac-LLY-AFC emits blue light (λmax = 400 nm); upon cleavage of the substrate by calpain, free AFC emits a yellow-green fluorescence (λmax = 505 nm), which can be quantified using a fluorometer or a fluorescence.