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 ContributorsPrefaceContents of Volume IAbbreviations List1 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 References2 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 References3 Two-Dimensional Gel Electrophoresis I. Introduction II. One-Dimensional Sodium Dodecyl Sulfate-Polyacrylamide Gels (SDS-PAGE) III. Two-Dimensional Gel Techniques References4 Measurements of Antibodies Specific for DNA I. Introduction II. Antigen Preparations III. Anti-DNA Assays IV. Critical Appraisal References5 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 References6 Methods in Surface Physics for Immunology I. Introduction II. Review of Surface Physics III. Methods of Measurement IV. Thin Layer Immunoassay Techniques V. Conclusions References7 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 References8 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 References9 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 References10 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 References11 A Helper Cell Assay of Cellular Antigens and Its Applications to Hapten-Specific ? Cells I. Introduction II. Principles of the Method and Its Modifications III. Materials IV. Procedures V. Comments References12 Limiting Dilution Analysis of Precursors of Cytotoxic ? 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 References13 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 References14 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 References15 Long-Term Culture and Cloning of Specific Helper ? Cells I. Objective and Principle of the Method II. Materials III. Procedure IV. Critical Appraisal Reference16 The Cloning of Alloreactive ? Cells I. Objective II. Principle III. Materials IV. Procedures V. Critical Appraisal References17 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 References18 Enzyme Immunoassay for the Detection of Hybridoma Products I. Introduction II. Materials III. Enzyme Immunoassay IV. Conclusion ReferencesIndex