Table of Contents

1. Introduction to Protein Purification Strategies 2. Preparation of Cellular and Subcellular Extracts Section 1: Purification Using Gel Electrophoresis 3. One-dimensional Polyacrylamide Gel Electrophoresis 4. Preparative Two-dimensional Gel Electrophoresis with Immobilized pH Gradients Section 2: Purification Using Column Chromatography 5. Ion-exchange Chromatography 6. Size-exclusion Chromatography 7. Reversed-phase High-performance Liquid Chromatography 8. Affinity and Immunoaffinity Chromatography 9. Metal Chelate Affinity Chromatography 10. Multidimensional Chromatography of Intact Proteins Section 3: Characterizing Protein Complexes 11. Identification of Protein-Protein Interactions by Conventional Column Chromatography 12. Identification of Associated Proteins by Coimmunoprecipitation 13. Chromatin Immunoprecipitation of Protein Complexes 14. Identification of Protein-Protein Interactions with Glutathione-S-Transferase Fusion Proteins 15. Direct Visualization of Protein Interactions in Living Cells Using Bimolecular Fluorescence Complementation Analysis 16. In Silico Tools: Analysis for Creating Focused Interaction Networks Appendices 1. Measuring the Concentration of Proteins 2. Concentrating Solutions of Proteins 3. Stabilization of Proteins for Storage 4. Immunoblotting and Staining of Membrane-bound Proteins 5. General Cautions Index