The Proteomics Protocols Handbook

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.

In The Protein Protocols Handbook, I have attempted to provide a cross-section of analytical techniques commonly used for proteins and peptides, thus providing a benehtop manual and guide both for those who are new to the protein chemistry laboratory and for those more established workers who wish to use a technique for the first time. We each, of course, have our own favorite, commonly used gel system, g- staining method, blotting method, and so on; I'm sure you will find yours here. H- ever, I have also described a variety of altematives for many of these techniques; though they may not be superior to the methods you commonly use, they may nev- theless be more appropriate in a particular situation. Only by knowing the range of techniques that are available to you, and the strengths and limitations of these te- niques, will you be able to choose the method that best suits your purpose.

This third volume provides comprehensive protocols on pre-analytical, analytical, plasma, and serum proteomics. New and updated chapters are divided into nine sections, detailing blood processing and handling strategies, discovery- and targeted-based mass spectrometry, including workflows to aid in discovery and targeted data analysis, in addition to software and bioinformatics for the plasma proteome. This edition further integrates emerging areas in the development of technologies for plasma proteomics and assay platforms in biomarker discovery and translational proteomics, enrichment and detection strategies to understand the plasma proteome, and peptide, lipid and metabolite targeted assays. We also detail the emerging analysis of extracellular vesicles isolated from plasma. Written in the format of the highly successful Methods in Molecular Biology series, each of the 33 chapters includes an introduction to the topic, lists necessary materials and methods, includes hints and tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Serum/Plasma Proteomics: Methods and Protocols, Third Edition aims to be comprehensive guide for researchers.

Shotgun Proteomics: Methods and Protocols serves as a vital collection of protocols through which thousands of proteins can be simultaneously identified, quantified, and characterized in a high throughput manner. Beginning with the history of proteomics centered on the vital role of mass spectrometry in its development, this detailed volume continues with chapters on sample pre-fractionation, in vivo and in vitro stable isotope labeling, label-free proteomics, informatics, protein-protein interactions, targeted proteomics, and post-translational modifications. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and comprehensive, Shotgun Proteomics: Methods and Protocols is an ideal and up-to-date guide for researchers seeking to understand the proteome of any given species.

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.

In this book, a select group of researchers has contributed their state-of-the-art methodologies on protein profiling and identification of disease biomarkers in tissues, microdissected cells and body fluids. The book integrates biochemistry, pathology, analytical technology, bioinformatics, and proteome informatics. Experimental approaches are thoroughly detailed and explained through a step-by-step instructional format that ensures successful results.

This volume presents the latest developments of the main pillars of protein analysis, such as sample preparation, separation and characterization. The book begins by describing basic but important sample preparation protocols. It then goes on to describe more sophisticated procedures on enriching specific protein classes and concludes with detailed descriptions of integrated work-flows for comprehensive protein analysis and characterization. The authors of the individual chapters are renowned protein biochemists who have all set value to provide a detailed representation of their lab work. Throughout the chapters, these authors share important tips and tricks for a successful and reproducible employment of their protocols in other laboratories. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Proteomic Profiling: Methods and Protocols is the perfect guide for students of Biochemistry, Biomedicine, Biology, and Genomics and will be an invaluable source for the experienced, practicing scientists.