Two-dimensional gel electrophoresis of proteins : methods and applications

著者

• Celis, J. E. (Julio E.)
• Bravo, R. (Rodrigo)

書誌事項

Two-dimensional gel electrophoresis of proteins : methods and applications

edited by Julio E. Celis, Rodrigo Bravo

Academic Press, 1984

注記

Includes bibliographies and index

内容説明・目次

内容説明

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.

目次

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 (R) 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

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詳細情報

• NII書誌ID(NCID): BA07082277
• ISBN:
  • 012164720X
• LCCN: 83005022
• 出版国コード: us
• タイトル言語コード: eng
• 本文言語コード: eng
• 出版地: New York
• ページ数/冊数: xvi, 487 p.
• 大きさ: 24 cm
• 分類:
  • LCC : QP551
  • DC19 : 574.19/245
• 件名:
  • LCSH : Proteins -- Analysis
  • LCSH : Electrophoresis, Polyacrylamide gel