Contents

Preface

Chapter 1

Introducing Standardization to Gel Electrophoresis by Comparative Fluorescence Gel Electrophoresis (CoFGE)

Abstract

Commentary

Acknowledgments

References

Chapter 2

Discontinuous Polyacrylamide Gel Electrophoresis

Abstract

Introduction

1. The Properties of Polyacrylamide Gels

1.1. The Polymerization Initiated by a Chemical Free Radical-Generating System

1.2. The Polymerization Initiated by a Photochemical Free Radical-Generating System

1.3. The Network Regulation and Porosity Control of Polyacrylamide Gels

2. The Essential Principles of Discontinuous Polyacrylamide Gel Electrophoresis in Technology

2.1. The Basic Concept of Discontinuous Polyacrylamide Gel Electrophoresis

2.2. The Principle of Stacking and Concentrating Effects of Discontinuous Polyacrylamide Gel Electrophoresis

2.2.1. Sample Stacking and Concentrating in Stacking Gels

2.2.2. Elimination of the Stacking Effect in Separating/Resolving Gels

2.2.3. Sample Concentrating in the Stacking Process

2.3. The Representative Buffer Systems for Discontinuous Polyacrylamide Gel Electrophoresis

2.3.1. The Tris(HCl and Tris(Gly Buffer System for Alkalic pH Conditions

2.3.2. The Acetic Acid(KOH and Acetic Acid(Gly/(-Alanine Buffer Systems for Acidic pH Conditions

2.3.3. The Representative Buffer Systems for Neutral pH Conditions

2.3.4. The Concentration of Discontinuous Buffer Systems

3. The Buffer Systems Used for Discontinuous Polyacrylamide Gel Electrophoresis

3.1. The Buffer Systems for the Discontinuous PAGE Performed in Alkalic pH Situations

3.2. The Buffer Systems for the Discontinuous PAGE Performed in Neutral pH Situations

3.2.1. The Tris(HCl and Tris(Barbita Buffer Systems

3.2.2. The Imidazole(Acetic Acid and Imidazole(Tricine Buffer Systems

3.2.3. The Bis-Tris(HEPES(MES/Acetic Acid and Bis-Tris(Tricine Buffer Systems

3.3. The Buffer Systems for Discontinuous PAGE Performed in Acidic pH Situations

3.3.1. The Acetic Acid(KOH and Acetic Acid((-Alanine/Gly Buffer Systems

3.3.2. The Citric Acid(Trisodium Citrate and Acetic Acid(Gly/(-Alanine Buffer Systems

3.3.3. The Acetic Acid(Sodium Acetate and Acetic Acid((-Alanine Buffer Systems

3.3.4. The Bis-Tris(HEPES(MES/Acetic Acid and Acetic Acid(6-Aminohexanoic Acid Buffer Systems

3. The Applications of Discontinuous PAGE Systems in Multiple Dimensional Electrophoresis

References

Appendix 1. The Buffer Systems Used for the Discontinuous PAGE Performed in Alkalic pH Situations

1.1. The Tris-HCl and Tris-Gly Buffer Systems

Appendix 2. The Buffer Systems for the Discontinuous PAGE Performed in Neutral pH Situations

2.1. The Tris(HCl and Tris(Barbital Buffer Systems

2.2. The Imidazole(Acetic acid and Imidazole(Tricine Buffer Systems

2.3. The Bis-Tris(HEPES(MES/Acetic Acid and Bis-Tris(Tricine Buffer Systems

Appendix 3. The Buffer Systems for the Discontinuous PAGE Performed in Acidic pH Situations

3.1. The Acetic Acid(KOH and Acetic Acid((-Alanine Buffer Systems

3.2. The Citric Acid(Trisodium Citrate and Acetic Acid(Gly/ (-Alanine Buffer Systems

3.3. The Acetic Acid(Sodium Acetate and Acetic Acid((-Alanine Buffer Systems

3.4. The Bis-Tris(HEPES(MES/Acetic Acid and Acetic Acid(6-Aminohexanoic Acid Buffer Systems

Chapter 3

Protein Electrophoresis in Avian Medicine

Abstract

Introduction

Serum or Plasma Proteins

Separation of Blood Proteins

Normal Protein Pattern

Alterations in Protein Pattern Caused by Non-Pathological Conditions

Protein Pattern Abnormalities

Protein Electrophoresis in Avian Clinical Practice

Conclusion

Acknowledgments

References

Chapter 4

Constant Field Gel Electrophoresis: Sensitive Method for DSBs Detection

Abstract

1. Introduction

1.1. DNA Damage

1.1.1. DSBs

1.1.2. Sources of DSBs DNA Damage

1.1.2.1. Endogenous

1.1.2.2. Exogenous

Ionizing Radiation (IR)

Ultraviolet radiation (UV)

DNA-damaging drugs

1.2. DSBs Repair Pathways

2. Evaluation of DSBs

2.1. Physical Approaches for DSBs Measurement

2.2. Biochemical Approaches for DSBs Measurement

2.2.1. TUNEL Technique

2.2.2. Flow cytometric (FCM) methods

2.2.3. Immunocytological Detection of DSBs

2.2.3.1. γ-H2AX Immunofluorescence Microscopy

2.2.4. Electrophoretic Methods

2.2.4 1. Comet Assay (SCGE)

2.2.4.2. Pulsed Field Gel Electrophoresis (PFGE)

2.2.4.3. Constant-Field Gel Electrophoresis (CFGE)

CFGE progress

Advantages

Disadvantages

Applications of CFGE

CFGE procedure

Parameters affecting DNA mobility and separation

Electric field strength

Cell (DNA) concentration in agarose plugs

Agarose concentration in gel

Electrophoresis buffers

Total electrophoresis duration (running time)

Bands staining

References

Biographical Sketches

Index

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