Immunological Methods
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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
Product details
- Language: English
- Copyright: © Academic Press 1981
- Published: May 28, 1981
- Imprint: Academic Press
- eBook ISBN: 9780323140058