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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.
List of contents
Two-Dimensional Gel Electrophoresis and 2D-DIGE.- Top-down Proteomics and Comparative 2D-DIGE Analysis.- DIGE Analysis Software and Protein Identification Approaches.- Native DIGE for Quantitative and Functional Analysis of Protein Interactomes.- Comparative Two-dimensional Fluorescence Gel Electrophoresis.- Identification of Ubiquitination associated Proteins using 2D-DIGE.- DIGE-based Phosphoproteomic Analysis.- DIGE-based Biomarker Discovery in Blood Cancers.- DIGE Saturation Labeling for Scarce Amounts of Protein from Formalin-fixed Paraffin-embedded (FFPE) Tissue.- DIGE Analysis of ProteoMiner(TM) Fractionated Serum/Plasma Samples.- Comparative 3-sample 2D-DIGE Analysis of Skeletal Muscles.- Proteomic identification of saliva proteins as non-invasive diagnostic biomarkers.- Analysis of the saliva proteome using 2D-DIGE.- DIGE Analysis of Clinical Specimens.- DIGE Analysis of Animal Tissues.- Rapid 2D-DIGE Proteomic Analysis of Mouse Liver within One Week.- 2D-DIGE analysis of liver disease in mice.- 2-Dye Versus 3-Dye DIGE for Comparative Testis Tissue Proteomic Analysis.- 2-CyDye based 2D-DIGE Analysis of Aged Human Muscle Biopsy Specimens.- Identification of Subproteomic Markers for Skeletal Muscle Profiling.- DIGE Analysis of Fish Tissues.- Sample preparation and protein determination for 2D-DIGE proteomics.- Protein Digestion for 2D-DIGE Analysis.- Subcellular Fractionation for DIGE-based Proteomics.- DIGE Analysis of Immunodepleted Plasma.- Bioinformatic Analysis of the Subproteomic Profile of Cardiomyopathic Tissue.- Elucidating Cellular Metabolism and Protein Difference Data from DIGE Proteomics Experiments using Enzyme Assays.- Enzyme Assay Methods to Validate DIGE Proteomics Data.- Immunoblot Analysis of DIGE-based Proteomics.- Verification of Protein Changes determined by 2D-DIGE based Proteomics using Immunofluorescence Microscopy.- Histological and Histochemical Microscopy used to verify 2D-DIGE Pathoproteomics.
