Caspase family of proteases are the central mediators of apoptosis in mammalian cells. The Caspase-12 Fluorometric BioAssay Kit provides a simple and convenient means for assaying the activity of caspases that recognize the sequence ATAD.Sample Type:Cell and tissue lysates Intended Use:For use in detecting the early/middle stages of apoptosis and differentiating apoptosis from necrosis.Specificity:Recognizes Caspase-12. Species Crossreactivity: mammalianTest Principle:The assay is based on detection of cleavage of substrate ATAD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). ATAD-AFC emits blue light (lmax = 400nm); upon cleavage of the substrate by caspase-12 or related caspases, free AFC emits a yellow-green fluorescence (lmax = 505nm), 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-12 activity.Kit Components:Cell Lysis Buffer, 1x25ml 2X Reaction Buffer, 1x2ml ATAD-AFC Substrate, 1x125ul DTT, 1M, 1x100ulStorage 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.
