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 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 Tribio^TM Caspase-3 fluorometric Assay Kits provide a simple and convenient method for the activity analysis of caspases that recognize the sequence DEVD. The assay is based on the detection of the free AFC fluorescence after cleavage from the labeled substrate DEVD-AFC. The free AFC can be quantified using a fluorometer or a fluorescence microtiter plate reader at Ex400/Em505nm. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows the determination of the fold increase in caspase-3 activity.

Applications

• Apoptosis
• Drug screening
• Growth factors
• Cytotoxicity