Spectrophotometry and Spectrofluorimetry

About this book

Spectrophotometry and Spectrofluorimetry: A Practical Approach Second Edition was written with the intention to help the reader understand the background concepts and practical applications of spectrophotometry and spectrofluorimetry. Optical spectroscopy underpins the day to day operations of most laboratories in the chemical, biological and medical sciences and this edition contains substantially updated and new chapters addressing the principles of most of the more common applications such as: spectrophotometry, spectrophotometric assays, spectrofluorimetry, time resolved fluorescence and phosphorescence studies, circular dichrosim and pre-equilibrium spectroscopic techniques. In all chapters, the emphasis is placed upon the practical aspects, with protocols to guide readers through test experiments. Other chapters are included to introduce subjects that have traditionally depended upon spectroscopy such as basic enzyme kinetics, ligand binding, data handling and the more recently established interest in the study of protein and DNA stability. Finally, the concept of 'global analysis' is introduced to provide the reader with an insight into this method of utilizing the vast arrays of experimental data provided by current instrumentation.

Contents

Introduction to light absorption; visible and ultraviolet spectra; Fluorescence principles and measurement; Time resolved fluorescence and phosphorescence spectroscopy; An introduction to circular dichroism; Quantitative determination of equilibrium binding isotherms for the multiple ligand - macromolecule interactions using spectroscopic methods; Steady state kinetics; Spectrophotometric assays; Stopped flow spectroscopy; Stopped flow fluorescence; Stopped flow circular dichroism; Spectrophotometry and fluorimetry of cellular compartments and intracellular processes; Use of optical spectroscopic methods to study the thermodynamic stability of proteins; The use of spectroscopic techniques in the study of protein stability

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