Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. The Caspase-4 Colorimetric BioAssay Kit provides a simple and convenient means for assaying the activity of caspases that recognize the sequence LEVD.Sample Type:Cell and tissue lysatesIntended Use:For use in detecting the early/middle stages of apoptosis and differentiating apoptosis from necrosis.Specificity:Recognizes Caspase-4. Species Crossreactivity: mammalianTest Principle:The assay is based on spectrophotometric detection of the chromophore p-nitroanilide (pNA) after cleavage from the labeled substrate LEVD-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 LEVD-dependent Caspase activity.Kit Components:Cell Lysis Buffer, 1x100ml 2X Reaction Buffer, 1x16ml LEVD-pNA, 4mM, 2x0.5ml DTT, 1M, 1x0.4ml Dilution Buffer, 1x200mlStorage 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.
