Enzyme assays are one of the most frequently performed procedures in biochemistry and are routinely used to estimate the amount of enzyme present in a cell or tissue, to follow the purification of an enzyme, or to determine the kinetic parameters of a system. The range of techniques used to measure the rate of an enzyme-catalysed reaction is limited only by the nature of the chemical change and the ingenuity of the investigator. This book describes the design and execution of enzyme assays, covering both general principles and specific examples. Chapters contain experimental protocols, including photomeric, electrochemical, radiochemical, and HPLC techniques, along with methods for determining enzyme activity after gel electrophoresis. The theory underlying each method is outlined, together with a description of the instrumentation, sensitivity and sources of error. Also discussed are the techniques and problems associated with enzyme extraction, statistical analysis of kinetic data, and a chapter examining buffers and methods of protein estimation. This work will be valuable not only to biochemists, but to researchers in all areas of the life sciences.
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Michael J. Danson, both Senior Lecturers in Biochemistry, University of Bath.
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