Apoptosis plays a fundamental role in many normal biological processes as well as in several disease states. Apoptosis can be induced by various stimuli that all produce the same end result: systematic and orderly cell death. Caspases cleave a variety of cellular substrates after aspartic acid residues—a characteristic that is central to their role in mammalian apoptosis. Caspases are synthesized in the cytosol of mammalian cells as inactive zymogens, which become active through intracellular caspase cascades. The TriboTM Caspase-3 Colormetric Assay Kits provides a simple and convenient means for assaying the activity of caspases that recognize the sequence DEVD. The assay is based on spectrophotometric detection of the chromophore pnitroaniline (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 caspase-3 activity.

Method:

Colorimetric

Storage Conditions:

Store Cell Lysis buffer and Assay buffer at 4℃, Store all other components at -20℃. Shelf life of six months.