Methods of Protein Separation (Paperback, Softcover Repri)

1 Sedimentation and Gel-Permeation Chromatography of Associating-Dissociating Macromolecules: The Role of Ligand Mediation and Rates of Reaction.- I. Introduction.- II. Theoretical Formulation.- III. Results.- A. Ligand-Mediated Association-Dissociation Reactions.- B. Kinetically Controlled Nonmediated Interactions.- IV. Discussion.- V. References.- 2 Trans Electrophoresis.- I. Introduction.- A. Principle.- B. History of in Situ Electro-Optical Scanning.- C. Types of Electrophoresis and Supporting Media.- D. Anatomy of the TRANS Electrophoresis System.- II. Instrumental Aspects.- A. The Electro-Optical Unit.- B. The Scanning Stage Module.- C. The Scanner Control Servo Unit.- D. The Electrophoresis Cell Cassette.- E. The Filling/Purging/Cooling Module.- F. Other Components.- G. The Digital Data Acquisition Module.- H. General Operational Procedure.- I. Column-Coating Procedure.- III. Data Processing.- A. Slope Analysis.- B. Moment Analysis.- IV. TRANS-CZE and TRANS-MZE.- A. Preliminary Considerations.- B. Peak Velocity.- C. Kinetic Peak-Variance Measurements.- D. Resolution.- E. Boundary Displacement.- V. TRANS-IF.- A. Preliminary Considerations.- B. Minimal Focusing Time.- C. Segmental pH Gradient and Apparent Isoelectric Point.- D. Resolving Power and Resolution.- E. Retardation Coefficient.- F. Kinetics of Defocusing and Refocusing.- G. Nonideal Effects in TRANS-IF.- VI. Future Developments.- VII. References.- 3 Immunodiffusion.- I. Introduction.- II. Antigen-Antibody Reactions.- III. Immunodiffusion.- IV. Immunodiffusion Combined with Electrophoresis.- V. Helpful Hints.- VI. References.- 4 Isoelectric Focusing in Polyacrylamide Gel.- I. Introduction.- II. Background.- III. Methodology.- A. Apparatus.- B. Electrolyte Solutions.- C. Gel Composition.- D. Electrolysis Conditions.- E. Sample Application.- F. Sample Load.- G. Measurement of pH Gradients.- H. pH-Gradient Instability.- IV. Sample Detection.- A. Staining for Proteins.- B. Histochemical Staining.- C. Gel Fractionation.- D. Recovery of Focused Zones.- E. Markers.- V. Applications.- A. Ferritins.- B. Hemoglobins.- C. Interacting Systems.- D. Nucleic Acids.- E. Two-Dimensional Procedures.- VI. Discussion.- VII. References.- 5 Purification of Chemically Modified Proteins.- I. Introduction.- II. General Types and Purposes of Chemical Modification.- A. Modification to Change Biological Activity.- B. Modification to Change Physical Properties.- C. Modifications to Block Deteriorative Reactions.- D. Introduction of a Label.- E. Reversible Modifications.- III. Some Problems Encountered in Chemical Modification.- A. Denaturation.- B. Reversibility of Modification.- C. Heterogeneity of Products.- D. A Product with Properties Very Similar to Those of the Original Native Protein.- E. Side Reactions of a Chemical Nature Occurring During Modification.- IV. Analysis of Chemically Modified Proteins.- A. Analysis of Amino Acid Residues.- V. Typical Examples of the Purification of Chemically Modified Proteins.- A. Heterogeneity Commonly Encountered in the Introduction of Groups Causing Changes in Charges.- B. Modifications with No Change in Charge.- C. Some Special Procedures for Chromatography.- VI. The Use of Chemical Modification as a Tool for Purification of Proteins.- A. Purification by Reversible-Complex Formation.- B. Reversible Chemical Modification for Separating Hybrids of Variants of Oligomeric Proteins.- C. The Use of Chemical Modification to Separate Products with Different Biochemical Activities.- D. Reversible Modification to Allow for Another Modification.- E. The Use of Cross-Linking Agents to Determine Degree of Polymerization and Localization of Proteins in Tissues.- VII. References.- 6 Chromatographic Peak Shape Analysis.- I. Introduction.- II. Moment Analysis.- A. Definition.- B. Mass Balance and the Moments.- C. Other Uses of Moments Analysis.- III. Experimental Studies.- A. Verification of Skew and Excess Utility.- B. Peak Identification.- IV. Slope Analysis.- V.