The second edition of this volume provides a comprehensive update of this key method on gel-based proteomics. Chapters present an introduction into the development of methods on principles of differential protein labeling and two-dimensional gel electrophoresis, techniques on optimized proteomic workflows using advanced mass spectrometry for protein identification, and the application of those methods in basic biological research, pathobiology and applied biomarker discovery. 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, Difference Gel Electrophoresis: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.