Two-Dimensional Gel Electrophoresis of Proteins

Methods and Applications

1st Edition - March 1, 1984
Editor: Julio Celis
Language: English
eBook ISBN:
9 7 8 - 0 - 3 2 3 - 1 4 7 9 1 - 0

Two-Dimensional Gel Electrophoresis of Proteins: Methods and Applications reviews current methods and clinical applications of two-dimensional gel electrophoresis of proteins,… Read more

Purchase options

LIMITED OFFER

Save 50% on book bundles

Immediately download your ebook while waiting for your print delivery. No promo code is needed.

Institutional subscription on ScienceDirect

Request a sales quote

Two-Dimensional Gel Electrophoresis of Proteins: Methods and Applications reviews current methods and clinical applications of two-dimensional gel electrophoresis of proteins, including the QUEST system, silver staining, and peptide mapping. Two-dimensional gel electrophoresis are applied to the study of diseases ranging from inborn errors of metabolism to human germ-line mutation rates, cancer, and mistranslation in animal and bacterial cells. This volume is organized into three sections encompassing 14 chapters and begins with an overview of the methodology of two-dimensional gel electrophoresis, followed by a discussion of computerized two-dimensional gel electrophoresis, silver staining, immunoblotting, and one- and two-dimensional peptide mapping. In most cases, a step-by-step guide to the techniques is given so that procedures may be easily repeated. A catalog of mouse fibroblast proteins is also given. The chapters that follow focus mainly on applications of two-dimensional gel electrophoresis in areas such as clinical and cancer research, human genetics, protein biosynthesis, and gene expression in plants. The final section presents current protein catalogs of Escherichia coli and human HeLa cells. This book is suitable for young researchers as well as for senior scientists working with a wide variety of problems in molecular and cell biology, basic biochemistry, genetics, and clinical research.

Contributors

Preface

Part I. Methods and Satellite Techniques

1. Two-Dimensional Gel Electrophoresis: A Guide for the Beginner

I. Introduction

II. Materials and Solutions

III. Experimental Procedures

IV. Hints and Comments

V. Concluding Remarks

References

2. The QUEST System for Computer-Analyzed Two-Dimensional Electrophoresis of Proteins

I. Introduction

II. Problems to Be Faced in Development of Computerized Protein Data Bases

III. Strategy for the QUEST System

IV. The QUEST System As Seen by the User

V. Program Descriptions

VI. Perspectives

References

3. Detection of Polypeptides in Two-Dimensional Gels Using Silver Staining

I. Introduction

II. Staining Procedures

III. Mechanisms of Silver Stains

IV. Potential and Real Problems in Silver Staining

References

4. Applicability of Color Silver Stain (GELCODE® System) to Protein Mapping with Two-Dimensional Gel Electrophoresis

I. Introduction

II. Variables that Affect Sensitivity and Reproducibility of the Color Silver Stain (GELCODE System)

III. Application of Color Silver Staining (GELCODE System) to Two-Dimensional Mapping of Proteins

IV. Computerized Data Acquisition of Color-Stained Images: Test for Stoichiometric Staining

V. Concluding Remarks

References

5. Electrophoretic Transfer of Proteins from Two-Dimensional Gels to Sheets and Their Detection

I. Introduction

II. Electrophoretic Transfer of Proteins

III. Probing the Transfer

IV. Removal of Probes

V. Examples and Applications of Protein Transfer Analysis

VI. Concluding Remarks

References

6. Correlation between Mouse and Human Two-Dimensional Gel Patterns: Peptide Mapping of Proteins Extracted from Two-Dimensional Gels

I. Introduction

II. Methods

III. Correlation between the Mouse and Human Two-Dimensional Gel Patterns

IV. Concluding Remarks

References

Part II. Applications of Two-Dimensional Gel Electrophoresis

7. Clinical Applications of Two-Dimensional Gel Electrophoresis

I. Introduction

II. The Electrophoresis System

III. Serum and Plasma

IV. Urine

V. Blood Cells

VI. Solid Tissue

VII. Other Specimens

VIII. Conclusions

References

8. Two-Dimensional Gel Electrophoresis for Studies of Inborn Errors of Metabolism

I. Introduction

II. Detection of Protein Polymorphism

III. Protein Mutation by Two-Dimensional Gel Electrophoresis

IV. Characteristic Patterns of Polypeptide Modulation

V. Conclusion

References

9. Adapting Two-Dimensional Gel Electrophoresis to the Study of Human Germ-Line Mutation Rates

I. Introduction

II. Some Basic Facts Concerning Mutation

III. Why Attempt to Use 2-D Gels to Study Mutation

IV. Some General Considerations in the Design of a Program to Detect Mutations

V. Selection of Most Appropriate Material for Study

VI. Technical Considerations

VII. A Statistical Analysis of Factors Influencing Spot Location

VIII. Ability of the System to Detect Genetic Variants

IX. Nomenclature

X. Gel Analysis

XI. Steps Necessary to Identify a Putative Mutant

XII. Data Management

XIII. The Selection of Populations for Study

XIV. Concluding Comments

References

10. Expression of Cellular Proteins in Normal and Transformed Human Cultured Cells and Tumors: Two-Dimensional Gel Electrophoresis as a Tool to Study Neoplastic Transformation and Cancer

I. Introduction

II. Transformation-Sensitive Polypeptides in Cultured Epithelial and Fibroblast Human Cells: Work in This Laboratory

III. Transformation-Sensitive Polypeptides of Known Identity

IV. Variation in the Expression of Mutant Actins in Chemically Transformed Human Diploid Fibroblasts: Actin in General

V. Phosphoproteins

VI. Polypeptide Synthesis in Human Tumors and Normal Tissue

VII. Conclusions

References

11. Application of Two-Dimensional Polyacrylamide Gel Electrophoresis to Studies of Mistranslation in Animal and Bacterial Cells

I. Introduction

II. Detection of Error-Containing Proteins by Two-Dimensional Polyacrylamide Gel Electrophoresis

III. Quantification of the Error Frequency of Protein Synthesis

IV. Assessment of the Error Frequencies of Protein Synthesis under Varying Physiological Conditions

V. Other Uses of the Two-Dimensional Gel Assay

VI. Two Out of Three Reading

References

12. Application of Two-Dimensional Gel Electrophoresis in Studies of Gene Expression during Early Plant Development

I. Introduction

II. Gene Expression during Early Embryogenesis

III. Somatic Embryogenesis

IV. Gene Expression during Somatic Embryogenesis

V. Coordinate Regulation of Sets of Biochemical Phenotypes

VI. Gene Expression in Temperature-Sensitive Cell Lines Impaired Embryogeny

VII. Gene Expression during Late Embryogeny

VIII. Gene Expression in Cotton Cotyledons

IX. Gene Expression in Germinating Seeds

X. Conclusion

References

Part III. Protein Catalogs

13. The Protein Catalog of Escherichia coli

I. Introduction

II. Identification of Proteins

III. Current E. coli Catalog

IV. Applications in E. coli Research

References

14. Catalog of HeLa Cell Proteins

I. Introduction

II. Two-Dimensional Gel Electrophoretic Separation of HeLa Cell Proteins

III. Some Characteristics of HeLa Cell Proteins

IV. Transformation-Sensitive Polypeptides

V. Conclusions

References

Index