The book deals with the theory and practice of all electrophoretic steps leading to proteome analysis, i.e. isoelectric focusing (including immobilized pH gradients), sodium dodecyl sulphate electrophoresis (SADS-PAGE) and finally two-dimensional maps. It is a reasoned collection of all modern, relevant, up-to-date methodologies leading to successful fractionation, analysis and characterization of every polypeptide spot in 2-D map analysis. It includes chapters on the most sophisticated mass spectrometry developments and it helps the reader in navigating through the most important databases in proteome analysis, including step by step tours in selected sites. Yet, this book's unique strength and feature is the fact that it combines not only practice (in common with any other book on this topic) but also theory, by giving a detailed treatment on the most advanced theoretical treatments of steady-state techniques, such as isoelectric focusing and immobilized pH gradients. A lot of this theory is newly developed and presented to the public for the first time. Thus, this book should satisfy not only the needs of every day practitioners, but also the desires of the most advanced theoreticians in the field, who will surely appreciate the novel theories presented here.
Also the methodological section contains several as yet unpublished protocols, correcting some of the existing ones and showing the pitfall and limitations of even well ingrained protocols in proteome analysis, which are here critically re-evaluated for the first time.
For researchers dealing with proteome analysis, and all those in working in the field of life sciences, medicine, pharmaceutics, and separation science.
Preface. Part I: Isoelectric Focusing: Fundamentals. Perspectives and Limits. Optimization of the Separation Process. Part I.I: Isoelectric Focusing: Fundamentals. Introduction. Isoelectric Focusing: Principles and Historical Aspects. References. Electrolyte Dissociation in Water Solution. Simple Electrolytes. Stepwise and parallel dissociation schemes for a bivalent protolyte. Relative concentration of different protolyte forms for stepwise and parallel schemes. Hydrogen ions concentration and buffer capacity. Ionization coefficient. Isoelectric point. Mobility of protolyte molecule. Non additive sum for buffer capacity in case of stepwise dissociation. Non amphoteric compounds and buffer capacity in "isoprotic state". References. Dissociation of Polyvalent Electrolytes. Acid-base equilibria, macroscopic and microscopic constants. Dissociation schemes of a hybrid type. Proton transfer tautomerism. Schemes with independent dissociation. Titration curve modeling. Linderstrøm-Lang equation. Calculation of the complete set of microconstants. Relative concentration of microstates for a homopolymer (independent dissociation). References. Kinetic Aspects of Acid-base Equilibria. Life-time of microscopic states. Relaxation of the ionic atmosphere. Modeling of the electrophoretic flux, electrophoretic mobility and conductivity. References. Natural pH Gradients. Simplest examples of natural pH-gradients. pH-gradients created with a multi-component mixture of amphoteric compounds. References. Immobilized pH Gradients. Classical immobilized pH-gradients created with linear density gradient. Linear pH-gra
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- © Elsevier Science 2001
- 25th October 2001
- Elsevier Science
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@from:D. Perrett @qu:...this is a valuable book that should be read by all those working in proteomics and interested in understanding the electrophoretic principles behind 'classical proteomics'. @source:Proteomics