Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. The Caspase-3/CPP32 Colorimetric BioAssay Kit provides a simple and convenient means for assaying the activity of caspases that recognize the sequence DEVD. Sample Type:Cell and tissue lysates Intended Use:For use in detecting the early/middle stages of apoptosis and to differentiate apoptosis from necrosis.Specificity:Recognizes Caspase-3. Species Crossreactivity: mammalianTest Principle:The assay is based on spectrophotometric detection of the chromophore p-nitroaniline (pNA) after cleavage from the labeled substrate DEVD-pNA. The pNA light emission can be quantified using a spectrophotometer or a microtiter plate reader at 400 or 405 nm. Comparison of the absorbance of pNA from an apoptotic sample with an uninduced control allows determination of the fold increase in CPP32 activity.Kit Components:Cell Lysis buffer, 1x100ml 2X Reaction Buffer, 4x2ml DEVD-pNA, 4mM, 1x0.5ml DTT, 1M, 1x0.4ml Dilution Buffer, 1x100mlStorage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
