Caspase-9 Fluorometric Assay Kit

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-9 is a member of the caspase family. Caspase-9, as well as caspase-2, -8 and -10, is an initiator caspase. Caspase-9 is involved in the mitochondrial death pathway and is activated during apoptosis. The activation of JNK/SAPK stress signaling pathways induces the release of cytochrome c from mitochondria and activation of apaf-1, which then cleaves caspase-9 into the active form. The active Caspase-9 cleaves and activates Caspase-3 and Caspase-7, which then cleave poly ADP ribose polymerase.

 Caspase-9 Fluorometric Assay Kit provides a convenient and simple way for detecting the LEHD-dependent caspase activity. LEHD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin) emits blue light (λmax = 400 nm); while cleavage of the substrate LEHD-AFC by Caspase-9 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 free AFC from an apoptotic sample with an uninduced control determines the fold increase in Caspase-9 activity.