Caspase-4 Assay Kit, Fluorometric with Ac-LEVD-AFC substrate

Caspase-4 Assay Kit, Fluorometricwith Ac-LEVD-AFC substrate

General description

Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. The Caspase-4 Fluorometric Assay Kit provides a simple and convenient means for assaying the activity of caspases that recognize the sequence LEVD. The assay is based on detection of cleavage of substrate LEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). LEVD-AFC emits blue light (400 nm); upon cleavage of the substrate by Caspase-4 or related caspases, free AFC emits a yellow-green fluorescence (505 nm), which can be quantified using a fluorometer or a fluorescence microtiter plate reader.

Detection Method:         Fluorescence (Ex/Em 400/505 nm)

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 only ~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 LEVD-dependent caspase activity

Kit Components :

• Cell Lysis Buffer
• 2X Reaction Buffer
• LEVD-AFC (1 mM)
• DTT (1 M)