Apoptosis is involved in a variety of physiological and pathological events, ranging from normal fetal development to development of diseases, such as cancer, organ failure and neurodegenerative diseases. Caspases are a large family of cysteinyl aspartate-specific proteases. They are activated in response to diverse cell death stimuli and ultimately dismantle a cell through restricted proteolysis of numerous cellular proteins, leading to apoptotic cell death. The SensoLyte® 520 Generic Caspase Assay Kit provides a method for in situ detection of caspase activity in cells undergoing apoptosis using a fluorochrome inhibitor of caspase, FAM-VAD-FMK. This reagent is cell permeable and non-cytotoxic. When added to a population of cells, it enters cells and covalently binds to activated caspases-1, -3 -4, -5, -7, -8, and –9. Bound FAM-VAD-FMK is retained within the cell, while any unbound FAM-VAD-FMK diffuses out of the cells and is washed away. Signal in the cells containing the bound FAM-VAD-FMK can be evaluated with a fluorescence plate reader, fluorescence microscopy, or flow cytometry.
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