Isoelectric Focusing

Isoelectric Focusing

1st Edition - January 1, 1976

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  • Editor: Nicholas Catsimpoolas
  • eBook ISBN: 9780323158053

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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.

Table of Contents

  • List of Contributors


    1 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


    2 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


    3 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


    4 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


    5 Isoelectric Focusing/Electrophoresis in Gels

    I. Introduction

    II. Aims of Gel Electrofocusing/Gel Electrophoresis

    III. General Techniques and Apparatus

    IV. Specific Combinations

    V. Conclusion


    6 Isoelectric Focusing in Granulated Gels

    I. Introduction

    II. Thin-Layer Isoelectric Focusing

    III. Preparative Isoelectric Focusing

    IV. Applications

    V. Technical Appendix


    7 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


    8 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 Μicroheterogeneity of Proteins


    9 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



Product details

  • No. of pages: 278
  • Language: English
  • Copyright: © Academic Press 1976
  • Published: January 1, 1976
  • Imprint: Academic Press
  • eBook ISBN: 9780323158053

About the Editor

Nicholas Catsimpoolas

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