Caspase-3 Fluorometric Assay Kit

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Background

Cysteine-dependent aspartate-directed proteases (Caspases) are a family of cysteine proteases that play important roles in apoptosis, necrosis, and inflammation. Sequential activation of caspases plays an important role in cell apoptosis. Caspase-3 is a caspase protein that cleaves and activates caspases 6 and 7, and is processed and activated by caspases 8, 9, and 10. Caspase-3 is the predominant caspase involved in the cleavage of amyloid-beta 4A precursor protein, which is associated with Alzheimer's disease. Caspase-3 recognizes tetra-peptide sequences D-x-x-D and hydrolyzes peptide bonds after aspartic acid residues.
Caspase-3 Fluorometric Assay Kit provides a convenient and simple way for detecting the DEVD-dependent caspase activity. DEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin) emits blue light (λmax = 400 nm); while cleavage of DEVD-AFC by Caspase-3 or related caspases, free AFC emits a yellow-green fluorescence (λmax = 505 nm), which can be quantified by using a fluorecence microtiter plate reader or a fluorometer. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control determines the fold increase in caspase-3 activity.