Immunological Methods

Volume IV

1st Edition - November 28, 1990
Imprint: Academic Press
Editors: Ivan Lefkovits, Benvenuto Pernis
Language: English
Paperback ISBN:
9 7 8 - 1 - 4 8 3 2 - 4 2 5 3 - 8
eBook ISBN:
9 7 8 - 1 - 4 8 3 2 - 6 5 3 6 - 0

Immunological Methods, Volume IV provides information pertinent to the methods in immunological research. This book focuses on cells, clones, and cell lines, as well as on their… Read more

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Immunological Methods, Volume IV provides information pertinent to the methods in immunological research. This book focuses on cells, clones, and cell lines, as well as on their components and secreted products. Organized into 21 chapters, this volume begins with an overview of hybridoma methodology as the most celebrated immunological method. This text then discusses cell fusion, hybridoma technology, and everything related to monoclonal antibodies. Other chapters consider another molecular biology method, which describes the procedure required for establishing a partitioned cDNA-library. This book provides as well a comprehensive analysis of mRNA populations in which every messenger species appears as a distinct element, and so provides accurate answers to questions concerning genetic complexity. The final chapter provides an example of how transgenic mice can be used to study the development of T cell repertoires. This book is a valuable resource for cell biologists, scientists, immunologists, and research workers.

Contributors

Preface

Contents of Previous Volumes

Abbreviations List

1 Solubilization of Cell Surface Membrane-Associated Molecules by Genetic Engineering

I. Introduction

II. General Strategy

III. Materials and Stock Solutions

IV. Experimental Procedures

V. Production of Solubilized Chimeric Molecules

References

2 Methods and Vectors for Positive Selection and Cell-Free Expression of Lymphocyte cDNA Clones

I. Introduction

II. Vector Design

III. Methods

IV. Critical Appraisal

References

3 Application of Polymerase Chain Reaction in Molecular Immunology

I. Polymerase Chain Reaction

II. General PCR Procedure

III. Applications

IV. Critical Appraisal

References

4 Target Detection Assay: A General Method to Determine DNA Binding Sites for Putative DNA-Binding Proteins

I. DNA-Protein Interactions

II. Principle of the Target Detection Assay

III. General Procedure to Determine DNA Target Sites by Target Detection Assay

IV. Applications

V. Critical Appraisal

References

5 Analysis of cis- and trans-Acting Factors Regulating Gene Transcription

I. Introduction

II. Transfection of Lymphoid Cells

III. Mobility Shift Assay

IV. Protein Footprinting

V. Protein Identification

VI. Purification of DNA-Binding Proteins by Affinity Chromatography

References

6 Cell-Free System for Polyadenylation Using Mouse B Cell Extracts

I. Introduction

II. Procedure

III. Critical Remarks

References

7 In Vitro Transcriptional Rate Assay for Lymphoid Cells

I. Introduction

II. Materials

III. Methods

IV. Applications

V. Critical Appraisal

References

8 System for High-Level Production in Escherichia coli and Rapid Purification of Recombinant Proteins: Application to Epitope Mapping, Preparation of Antibodies, and Structure-Function Analysis

I. Introduction

II. Expression of the Recombinant Proteins in E. coli

III. Purification of Recombinant Proteins by Nickel Chelate Affinity Chromatography

IV. Application 1: Definition of Novel B Cell Epitopes in the Circumsporozoite Protein of Plasmodium falciparum

V. Application 2: Mapping of Functional Epitopes of Human IFN-γ Receptor

VI. Conclusions

VII. Appendix

References

9 Analysis of Two-Dimensional Gel Electrophoretic Protein Patterns upon in Vivo Labeling of Mice

I. Objective

II. Experimental Procedures

III. Applications

References

10 Biological Assay of Interleukins

I. Introduction

II. Immunoassays

III. Bioassays

IV. Critical Appraisal

References

11 Monoclonal Antibodies Against Cytokine Receptors

I. Introduction

II. Radioiodination of the Ligand

III. Assay of Cell Extracts for Ligand Binding

IV. Isolation of the Receptor Protein

V. Immunization

VI. Screening for Antireceptor Antibodies

VII. Appraisal

References

12 Human Monoclonal Antibodies

I. Introduction

II. Materials

III. Source and Purification of Human Lymphocytes

IV. Fusion Partners

V. In Vitro Immunization

VI. Fusion Conditions

VII. Purification of Human Monoclonal Antibodies

VIII. Concluding Remarks

References

13 Isolation of Rat Immunoglobulin Class Switch Variants from Hybridomas by Sequential Sublining

I. Introduction

II. Experimental Methods

III. Applications

References

14 Mapping of T Cell Epitopes in Proteins: A Chemical Approach

I. Objective and Principle of the Method

II. Experimental Procedure

References

15 Plasmodium Falciparum Malaria Parasite Cultures and Their Use in Immunology

I. Introduction

II. Materials

III. Methods

References

16 A Chemotactic Assay Enabling Recovery of Cells after Migration

I. Introduction

II. Analytical Chambers

III. Preparative Chambers

References

17 Assay for Quantitation of Cell-Cell Adhesion Using Fluorescent Dyes

I. Introduction

II. Choice of a Fluorescent Cell Marker

III. Adhesion Assay

IV. Applications

References

18 Analysis of Avian T and B Lymphocytes

I. Introduction

II. Monoclonal Antibodies

III. Analysis of T and B Cells

References

19 Isolation of Cells Involved in the Germinal Center Reaction: Germinal Center B Cells and Follicular Dendritic Cells

I. Introduction

II. Principle of the Methods

III. Materials

IV. Procedure

V. Critical Factors and Comments

References

20 Applications of Fetal Thymus Organ Cultures in Studies of T Cell Development

I. Introduction

II. Materials

III. Methods

IV. Applications

References

21 Transgenic Mice for Analysis of T Cell Development

I. Introduction

II. Generation of Transgenic Mice

III. Analysis of Transgenic Mice

IV. Outlook

References

Index

Life Sciences

Physical Sciences & Engineering

Social Sciences & Humanities

Health

Immunological Methods

Volume IV

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Ivan Lefkovits

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