Proteome Research: Two-Dimensional Gel Electrophoresis and Identification Methods

摘要: 1 Introduction: The Virtue of Proteomics.- 2 Solubilization Proteins in 2D Electrophoresis.- Introduction.- Rationale Solubilization-Breaking Molecular Interactions.- 3 Initial Solubilization.- 4 Solubility During IEF.- 5 Conclusions: Current Limits and How to Push Them.- References.- Two-Dimensional Electrophoresis with Carrier Ampholytes.- Sample Preparation.- First Dimension. Standard Isoelectric Focusing.- Second Slab Gel Modification the Method.- 6 Visualization Separated Proteins: Silver Staining.- Appendix: Problems Troubleshooting.- Immobilized pH Gradients.- Dimension: IEF IPGs.- Equilibration IPG Strips.- SDS-PAGE.- Appendix A: General B: Troubleshooting for Horizontal PAGE.- Detection on Gels.- by Organic Dyes.- Differential Precipitation Salts.- Metal Ion Reduction (Silver Staining).- Fluorescence.- Radioactive Isotopes.- 7 Conclusions Future Prospects.- Blotting Immunoaffinity Identification Electrophoresis-Separated Proteins.- Protein Transfer onto Membranes.- Membrane Immunodetection.- Assignment Immunoreactive Spots Matching.- Amino Acid Composition After Hydrolysis.- from Gels PVDF Hydrolysis PVDF-bound Extraction Acids Derivatization Chromatography.- Analysis Guide.- Database 8 N-Terminal Sequence Tags Composition.- 9 Conclusions.- Obtained Labeling.- Choice Labeling Acids.- Single Double Method Based 35S Decay.- using Scintillation Counting.- Determination pI Mr.- Construction Database.- Search a Result Analysis.- 10 Conclusion.- Sequencing.- Purpose Edman Chemistry.- Other Possibilities Generation Sequences.- Interface Gel.- Perspectives.- Mass Spectrometry.- Preparation Methods Compatible MS.- Enzymatic Digestion Clean-up ESI-MS On-line Liquid MALDI-MS.- Spectrometers.- Correlating MS DATA Databases.- 11 Spectrometry Intact Elution Electroblotted Polymer UV-MALDI Directly Polyacrylamide References.