Abstract
This chapter discusses flow cytometric analysis procedures to measure the dynamics of enzyme activity. Enzymes, in one form or another, perform the majority of dynamic molecular interactions responsible for function in the intact living cell. Measurement of such dynamic cellular events in intact cells under near-physiological conditions, irrespective of the assay system, is arguably one of the most powerful techniques in cell biology as physiological and pathological processes are not static but are continuously variable. Clearly, an understanding of the dynamics of enzyme activity will be of crucial importance in attempts to modulate pathological states. The chapter shows a schematic for the conversion of substrate to product by enzyme action within whole cells. A protocol for quantitating nonspecific esterase activity using flow cytometry in viable intact cells is given in the chapter.
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