Caspase-3, 7 Assay Kit, Fluorometric with Ac-DEVD-AFC substrate

Caspase-3, 7 Assay Kit, Fluorometric with Ac-DEVD-AFC substrate

General description

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 fluorescence 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:         Fluorescence (400 nm excitation filter and 505 nm emission filter)

Species Reactivity:        Mammalian

Applications:                  Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis

Storage temp.:               -20°C

Features & Benefits:

• Simple one-step procedure that 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)