Immunological Methods - 1st Edition - ISBN: 9780124427020, 9780323140058

Immunological Methods

1st Edition

Authors: Author Unknown
eBook ISBN: 9780323140058
Imprint: Academic Press
Published Date: 28th May 1981
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Description

Immunological Methods, Volume II compiles procedures that are appropriate for studies in immunology.

This book discusses the determination of equilibrium binding parameters of monoclonal antibodies specific for cell surface antigens; two-dimensional gel electrophoresis; and measurements of antibodies specific for DNA. The methods in surface physics for immunology; HLA-DR typing by complement-dependent B lymphocyte lysis; and protein A plaque assay for the detection of immunoglobulin-secreting cells are also elaborated.

This text also covers the in vitro production and testing of antigen-induced mediators of helper T-cell function; limiting dilution analysis of precursors of cytotoxic T lymphocytes; and induction of antibody formation in mouse bone marrow. Other topics include the long-term culture and cloning of specific helper T cells; cloning of alloreactive T cells; and enzyme immunoassay for the detection of hybridoma products.

This publication is valuable to immunologists and medical practitioners researching on immunological methods.

Table of Contents


List of Contributors

Preface

Contents of Volume I

Abbreviations List

1 Determination of Equilibrium Binding Parameters of Monoclonal Antibodies Specific for Cell Surface Antigens

I. Introduction

II. Theoretical Background

III. Technical Approach

IV. Examples

V. Critical Appraisal

VI. Appendix

References

2 Biochemical Characterization of Cell Surface Antigens Using Monoclonal Antibodies

I. Introduction

II. Production of Antibodies

III. Labeling Procedures for Cell Surface Antigens

IV. Preparation of Cell Membranes

V. Analysis of Cell Surface Antigens

VI. Conclusion

References

3 Two-Dimensional Gel Electrophoresis

I. Introduction

II. One-Dimensional Sodium Dodecyl Sulfate-Polyacrylamide Gels (SDS-PAGE)

III. Two-Dimensional Gel Techniques

References

4 Measurements of Antibodies Specific for DNA

I. Introduction

II. Antigen Preparations

III. Anti-DNA Assays

IV. Critical Appraisal

References

5 High-Pressure Liquid Chromatography of Proteins and Peptides

I. Introduction

II. General Description of HPLC

III. Components of an HPLC System

IV. Examples of HPLC Applications

References

6 Methods in Surface Physics for Immunology

I. Introduction

II. Review of Surface Physics

III. Methods of Measurement

IV. Thin Layer Immunoassay Techniques

V. Conclusions

References

7 Hapten-Modified Antibodies Specific for Cell Surface Antigens as a Tool in Cellular Immunology

I. Introduction

II. Hapten Sandwich Labeling

III. Preparation of Hapten Sandwich Labeling Reagents

IV. Selected Examples of the Application of Hapten Sandwich Labeling Reagents

V. Critical Appraisal

References

8 HLA-DR Typing by Complement-Dependent B Lymphocyte Lysis

I. Introduction

II. Principle of the Test

III. Details of the Test

IV. Modifications of the Test

V. Critical Appraisal

VI. Equipment, Supplies, and Solutions

References

9 The Protein A Plaque Assay for the Detection of Immunoglobulin-Secreting Cells

I. Introduction

II. Reagents

III. Cell Suspensions

IV. Plaquing Procedure

V. Counting of Plaque-Forming Cells

VI. General Comments

VII. Appendix

References

10 In Vitro Production and Testing of Antigen-Induced Mediators of Helper T-Cell Function

I. Introduction

II. Materials

III. Use of the Marbrook Culture System for Inducing Antigen-Specific Helper Supernatants

IV. Testing the Activity of Antigen-Specific Factors

V. Plaque Assay

VI. Summary

References

11 A Helper Cell Assay of Cellular Antigens and Its Applications to Hapten-Specific Τ Cells

I. Introduction

II. Principles of the Method and Its Modifications

III. Materials

IV. Procedures

V. Comments

References

12 Limiting Dilution Analysis of Precursors of Cytotoxic Τ Lymphocytes

I. Principle of the Method

II. Materials

III. Methods

IV. Experimental Design

V. Analysis of the Results

VI. Developments of the Limiting Dilution Assay

VII. Limitations of the Assay

References

13 Induction of an Antibody Response in Cultures of Human Peripheral Blood Lymphocytes

I. Objective

II. Principle of the Method

III. Materials

IV. Procedure

V. Comments

References

14 Induction of Antibody Formation in Mouse Bone Marrow

I. Introduction

II. Collection of Mouse Bone Marrow Cells

III. Distribution of Antibody-Forming Cells Over Various Bone Marrow Compartments

IV. Calculation of the Antibody-Forming Cell Activity of the Total Bone Marrow

V. Kinetics of the Response

VI. Critical Factors

References

15 Long-Term Culture and Cloning of Specific Helper Τ Cells

I. Objective and Principle of the Method

II. Materials

III. Procedure

IV. Critical Appraisal

Reference

16 The Cloning of Alloreactive Τ Cells

I. Objective

II. Principle

III. Materials

IV. Procedures

V. Critical Appraisal

References

17 The Technique of Hybridoma Production

I. Introduction

II. Prefusion Steps

III. Preparation of Medium Constituents, Media, and Cell Suspensions

IV. Cell Fusion with Polyethylene Glycol

V. Products of Fusion

VI. Freezing and Thawing of Hybridoma Cells

References

18 Enzyme Immunoassay for the Detection of Hybridoma Products

I. Introduction

II. Materials

III. Enzyme Immunoassay

IV. Conclusion

References

Index

Details

Language:
English
Copyright:
© Academic Press 1981
Published:
Imprint:
Academic Press
eBook ISBN:
9780323140058

About the Author

Author Unknown