Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. The CPP32/Caspase-3 Fluorometric Protease Assay Kit provides a simple and convenient means for assaying the DEVD-dependent caspase activity. The assay is based on detection of cleavage of substrate DEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). DEVD-AFC emits blue light (λmax = 400 nm); upon cleavage of the substrate by CPP32 or related caspases, free AFC emits a yellow-green fluorescence (lmax = 505 nm), which can be quantified using a fluorometer or a fluorecence microtiter plate reader. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-3/CPP32 activity.
|Detection Method||Fluorecence (400 nm excitation filter and 505 nm emission filter)|
|Features & Benefits||• Simple one-step procedure; takes 1-2 hours
• Fast and convenient
• Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-3/CPP32 activity
|Kit Components||• Cell Lysis Buffer
• 2X Reaction Buffer
• DEVD-AFC (1 mM)
• DTT (1 M)
20tests ; 50tests ; 100tests
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