Two-Dimensional Electrophoresis: An Overview -- Solubilization of Proteins in 2DE: An Outline -- Selection of pH Ranges in 2DE -- Difficult Proteins -- Organelle Proteomics -- Applications of Chemical Tagging Approaches in Combination with 2DE and Mass Spectrometry -- Immunoblotting 2DE Membranes -- Troubleshooting Image Analysis in 2DE -- Analysis of Bacterial Proteins by 2DE -- Proteomic Analysis of Caenorhabditis elegans -- Protein Extraction for 2DE -- Analysis of Proteins from Marine Molluscs -- Preparation and Analysis of Plant and Plastid Proteomes by 2DE. -- High-Resolution 2DE -- Blue Native-Gel Electrophoresis Proteomics -- 2DE for Proteome Analysis of Human Metaphase Chromosomes -- Microsomal Proteomics -- Prefractionation Using Microscale Solution IEF -- Diagonal Electrophoresis for Detection of Protein Disulphide Bridges -- High-Resolution Large-Gel 2DE -- Silver Staining of Proteins in 2DE Gels -- Detection of 4-Hydroxy-2-Nonenal- and 3-Nitrotyrosine-Modified Proteins Using a Proteomics Approach -- Proteomic Detection of Oxidized and Reduced Thiol Proteins in Cultured Cells -- Detection of Ubiquitination in 2DE -- Phosphoproteome Analysis by In-Gel Isoelectric Focusing and Tandem Mass Spectrometry -- Detection of Protein Glutathionylation -- Activity-Based Protein Profiling of Protein Tyrosine Phosphatases -- Active Protease Mapping in 2DE Gels -- Two-Dimensional Difference Gel Electrophoresis -- Protein Expression Profiling -- C-Terminal Sequence Analysis of 2DE-Separated Proteins -- Shotgun Protein Analysis by Liquid Chromatography-Tandem Mass Spectrometry -- de Novo Sequence Analysis of N-Terminal Sulfonated Peptides After in-Gel Guanidination -- Tryptic Digestion of In-Gel Proteins for Mass Spectrometry Analysis -- Database Interrogation Algorithms for Identification of Proteins in Proteomic Separations -- Creating 2DE Databases for the World Wide Web.

Two-dimensional electrophoresis (2-DE) remains one of the most popular techniques for proteomic comparisons. Revolutionized by the development of immobilized pH gradient strips, this vital process has benefited greatly from recent developments in staining, mass spectrometry, and bioinformatics. In Two-Dimensional Electrophoresis Protocols, expert researchers examine these cutting edge proteomics methodologies, explore their application to a wide range of biological materials, and look at how these developments have helped standardize the design, execution, and analysis of proteomics experiments. Chapters reflect key steps in 2-DE experiments, including sample preparation, staining, post-translational modification, spot identification, and bioinformatics. Composed in the highly successful Methods in Molecular Biology series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Innovative and reader-friendly, Two-Dimensional Electrophoresis Protocols encourages newcomers to apply powerful 2-DE techniques to their own research, while also providing current and essential information for seasoned scientists.