Provided in this book are guidelines and practical advice for anyone working with enzymes. It offers advice on: how solvents, salts, substrates, and denaturants affect enzyme function and stability; developing a scheme for purifying an enzyme from different kinds of tissue; applying different techniques for fractionating proteins - ammonium sulfate, polyethylene glycol, and organic solvent fractionation - ion exchange, dye ligand, and bio-ligand affinity chromatography; characterizing proteins by determining molecular weight, isoelectric pH, amino acid composition and sequence, ligand binding and enzyme kinetics. Basic guidelines to a procedure and hints for operation of a technique are offered along with many references to the literature which provides additional details and theory. Methods for treating electrophoretic, gel permeation, enzyme kinetic and binding data are treated in detail. Various protein quantitation methods are described and their sensitivities are compared.
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Provided in this book are guidelines and practical advice for anyone working with enzymes. It offers advice on: how solvents, salts, substrates, and denaturants affect enzyme function and stability; developing a scheme for purifying an enzyme from different kinds of tissue; applying different techniques for fractionating proteins - ammonium sulfate, polyethylene glycol, and organic solvent fractionation - ion exchange, dye ligand, and bio-ligand affinity chromatography; characterizing proteins by determining molecular weight, isoelectric pH, amino acid composition and sequence, ligand binding and enzyme kinetics. Basic guidelines to a procedure and hints for operation of a technique are offered along with many references to the literature which provides additional details and theory. Methods for treating electrophoretic, gel permeation, enzyme kinetic and binding data are treated in detail. Various protein quantitation methods are described and their sensitivities are compared.
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