Isoelectric focusing (IEF) is a high-resolution, stand-alone technique that can be used as an analytical method or tool for protein purification. The only current book on the market, the Handbook of Isoelectric Focusing and Proteomics is the ideal 'one-stop' source for germane information in this discipline. This highly practical book also contains chapters on alternative methods that may pave the way in the search for efficient techniques for fractionating and purifying proteins. Complete with the history of IEF focusing to authors' insights and practical tips, this book is a must for anyone working in proteomics.* Is the only current book available on the subject * Includes author insights and practical tips* Is an ideal single source for students and researchers working in proteomics

Isoelectric focusing (IEF) is a high-resolution, stand-alone technique that can be used as an analytical method or tool for protein purification. The only current book on the market, the Handbook of Isoelectric Focusing and Proteomics is the ideal 'one-stop' source for germane information in this discipline. This highly practical book also contains chapters on alternative methods that may pave the way in the search for efficient techniques for fractionating and purifying proteins. Complete with the history of IEF focusing to authors' insights and practical tips, this book is a must for anyone working in proteomics. * Is the only current book available on the subject * Includes author insights and practical tips * Is an ideal single source for students and researchers working in proteomics

Still the only concise practical guide to laboratory experiments in proteomics, this new edition now also covers DIGE technology and liquid-chromatography, while the troubleshooting section has been considerably extended. Adopting a practical approach, the authors present the relevant techniques and explain the route to successful experimental design and optimal method selection. They cover such electrophoretic techniques as isoelectric focusing, SDS page, 2-D page, and DIGE, as well as liquid-chromatography techniques, such as ion exchange, affinity chromatography and reversed-phase HPLC. Mass-spectrometric techniques include MALDI, ESI, and FT ICR. Generously illustrated, partly in color, the book also features updates of protocols as well as animations illustrating crucial methodological steps on a companion website.

Hands-on researchers describe in step-by-step detail 73 proven laboratory methods and bioinformatics tools essential for analysis of the proteome. These cutting-edge techniques address such important tasks as sample preparation, 2D-PAGE, gel staining, mass spectrometry, and post-translational modification. There are also readily reproducible methods for protein expression profiling, identifying protein-protein interactions, and protein chip technology, as well as a range of newly developed methodologies for determining the structure and function of a protein. The bioinformatics tools include those for analyzing 2D-GEL patterns, protein modeling, and protein identification. All laboratory-based protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

The third edition of The Protein Protocols Handbook introduces 57 critically important new chapters, and significantly updates the previous edition's tried-and-trusted methods. The book offers over 200 key, readily reproducible protocols that ensure results.

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.

Recent advances in the biosciences have led to a range of powerful new technologies, particularly nucleic acid, protein and cell-based methodologies. The most recent insights have come to affect how scientists investigate and define cellular processes at the molecular level. This book expands upon the techniques included in the first edition, providing theory, outlines of practical procedures, and applications for a range of techniques. Written by a well-established panel of research scientists, the book provides an up-to-date collection of methods used regularly in the authors’ own research programs.