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Isoelectric Focusing
- 1st Edition - January 1, 1976
- Editor: Nicholas Catsimpoolas
- Language: English
- Hardback ISBN:9 7 8 - 0 - 1 2 - 1 6 3 9 5 0 - 1
- Paperback ISBN:9 7 8 - 0 - 1 2 - 4 3 3 4 9 3 - 9
- eBook ISBN:9 7 8 - 0 - 3 2 3 - 1 5 8 0 5 - 3
Isoelectric Focusing deals with the theory and applications of isoelectric focusing. The book reviews some alternative approaches to generation of pH gradients for isoelectric… Read more
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Request a sales quoteIsoelectric 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 Contributors
Preface
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
References
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
References
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
References
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
References
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
References
6 Isoelectric Focusing in Granulated Gels
I. Introduction
II. Thin-Layer Isoelectric Focusing
III. Preparative Isoelectric Focusing
IV. Applications
V. Technical Appendix
References
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
References
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
References
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
References
Index
- No. of pages: 278
- Language: English
- Edition: 1
- Published: January 1, 1976
- Imprint: Academic Press
- Hardback ISBN: 9780121639501
- Paperback ISBN: 9780124334939
- eBook ISBN: 9780323158053
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