Isoelectric Focusing deals with the theory and applications of isoelectric focusing. The book reviews some alternative approaches to generation of pH gradients for isoelectric focusing analysis, along with other relevant topics such as the theoretical aspects of steady-state isoelectric focusing, gel electrofocusing/gel electrophoresis, and transient state isoelectric focusing. This volume is organized into nine chapters and begins by looking at the history of the discovery of isoelectric focusing and correcting some blurred impressions conveyed in the literature. It then turns to a discussion of the differential equation of isoelectric focusing, the carrier ampholytes used in carrying out the technique, the use of isoelectric focusing in polyacrylamide and granulated gels as well as in free solution, and continuous-flow electrophoresis and isoelectric focusing. An attempt is made to illuminate problems in isoelectric focusing and to point out possible solutions in the areas of theory, mechanisms, ampholyte chemistry, and apparatus. Some of the processes that occur during an isoelectric focusing experiment are also considered. This book is intended for anyone who needs to know what isoelectric focusing is all about.

List of ContributorsPreface1 Isoelectric Focusing I. An Essential Condition for Isoelectric Focusing II. Isoelectric Focusing-A Family of Three Effects III. Demonstration of the Focusing and Evacuation Effects IV. Limitations of the Prepared pH Gradient Method and the Success of the Natural pH Gradient Method V. Some Recent Alternative Approaches to Generation of pH Gradients for Isoelectric Focusing Analysis References2 Theoretical Aspects of Steady-State Isoelectric Focusing I. Introduction II. The Differential Equation of Isoelectric Focusing III. Convection-Free Electrolysis of a Single Strong Acid or Strong Base IV. Convection-Free Electrolysis of a Single Weak Acid or Weak Base V. Convection-Free Electrolysis of an Acidic or Basic Ampholyte with Only Two Protolytic Groups in the Isoelectric Region VI. Convection-Free Electrolysis of a Mixture of Two Ampholytes VII. Convection-Free Electrolysis of a Mixture of Many Ampholytes References3 The Carrier Ampholytes I. Desirable Properties II. Synthesis of a System of Carrier Ampholytes III. Physicochemical Properties: Buffer Capacity and Conductivity IV. Number of Carrier Ampholyte Species with Different pI V. Light Absorption VI. Determination of pi of Proteins in the Presence of Carrier Ampholytes and Other Additives VII. Stability of Proteins and Use of Additives VIII. Separation of Carrier Ampholytes from Proteins IX. Possible Interaction with Proteins X. Biological and Biochemical Properties XI. Other Aspects References4 Isoelectric Focusing on Polyacrylamide Gel I. Introduction: Isoelectric Focusing in Transition II. Theory of Isoelectric Focusing III. Interpretation of pI IV. Mechanism of Isoelectric Focusing V. Synthetic Carrier Ampholytes VI. Apparatus: Analytical and Preparative VII. Gel VIII. Protein Fractionation References5 Isoelectric Focusing/Electrophoresis in Gels I. Introduction II. Aims of Gel Electrofocusing/Gel Electrophoresis III. General Techniques and Apparatus IV. Specific Combinations V. Conclusion References6 Isoelectric Focusing in Granulated Gels I. Introduction II. Thin-Layer Isoelectric Focusing III. Preparative Isoelectric Focusing IV. Applications V. Technical Appendix References7 Continuous-Flow Isoelectric Focusing I. Introduction II. Scaling-Up of Electrophoretic Separations III. Continuous-Flow Electrophoresis IV. Advantages of Continuous-Flow Isoelectric Focusing V. Experimental Methods VI. Practical Considerations VII. Conclusions References8 Isoelectric Focusing in Free Solution I. Introduction II. Comparison of Different Methods of Isoelectric Focusing in Free Solution III. Zone Convection Isoelectric Focusing IV. Free Zone Electrophoresis V. Free Zone Isoelectric Focusing VI. Free Isoelectric Focusing VII. The ¿icroheterogeneity of Proteins References9 Transient State Isoelectric Focusing I. Introduction II. Instrumental Aspects III. Data Acquisition and Processing IV. Methodological Parameters V. Kinetics of Defocusing and Refocusing VI. Remarks ReferencesIndex