Caspase-9 Inhibitor Drug Screening Kit (Fluorometric)

Caspases (Cysteine-dependent aspartate-directed proteases) are a family of cysteine proteases that play important roles in apoptosis, inflammation and necrosis. Sequential activation of caspases plays an important role in cell apoptosis. Inhibition of caspases can delay apoptosis, indicating a potential role in drug screening efforts. 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.

 The Caspase-9 Inhibitor Drug Screening Kit (Fluorometric) provides a simple, fast and convenient way for screening of caspase-9 inhibitors based on fluorometric method. The synthetic peptide substrate LEHD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin) emits blue light (λmax = 400 nm). While cleavage of LEHD-AFC by active caspase-9, free AFC emits a yellow-green fluorescence (λmax = 505 nm) that can be quantified by a fluorecence microtiter plate reader or a fluorometer. Inhibitors can be directly added to the reaction and the efficacy of inhibition of caspase-9 activity is determined by comparison of the fluorescence intensity in samples without and with the testing inhibitors. The kit is suited to perform directly in microtiter plates.