Immunological Methods

Ivan Lefkovits, Benvenuto Pernis

• Publisher: Academic Press
• Year: 2014
• Print ISBN: 9780124427501
• eBook ISBN: 9781483269993
• Topic: Biological Sciences
• Subtopic: Zoology
• Index: Biological Sciences

Immunological Methods a compendium of basic research techniques being used in one of the largest immunology research institutes, the Basel Institute for Immunology, with particular emphasis given to new methodology. The procedures have been described by individuals judged to be highly expert in their specialties. In many instances the methods developed or adapted to unique uses by the contributors have not previously been described in detail. The book contains 34 chapters covering techniques for detection, isolation, and purification of antibodies (including dansylation, two-dimensional chromatography, isoelectric focusing, polyacrylamide gel electrophoresis, and isotachophoresis); measurement of equilibrium constants (equilibrium dialysis, filtration, and sedimentation); and isotope and fluorescent labeling and detection of cell-surface components. Techniques such as isotope laboratory maintenance; chemical modification of proteins, haptens, and solid supports, and haptenation of viable biological carriers; production of antisera against allotypes and histocompatibility antigens and production of antibody with clonai dominance; histocompatibility and MLR testing; and cell separation by haptenated gels and by velocity sedimentation of rosette-forming cells are also discussed. Other chapters cover detection of antibody-secreting and alloantigen-binding cells; immune responses in vitro and their analysis by limiting dilution; production of T-cell factors; hybridoma production by cell fusion; maintenance of cell lines and cloning in semisolid media; and the mathematical analysis of immunological data.

Table of contents

• Cover image
• Title page
• Table of Contents
• Copyright
• List of Contributors
• Preface
• Abbreviations List
• Chapter 1: The Quality of Antibodies and Cellular Receptors
• Chapter 2: The Isolation and Characterization of Immunoglobulins, Antibodies, and Their Constituent Polypeptide Chains
• Chapter 3: Peptide Mapping at the Nanomole Level
• Chapter 4: Electrophoresis of Proteins in Polyacrylamide Slab Gels
• Chapter 5: Resolution of Immunoglobulin Patterns by Analytical Isoelectric Focusing
• Chapter 6: Isolation of Monoclonal Antibody by Preparative Isoelectric Focusing in Horizontal Layers of Sephadex G-75
• Chapter 7: Isotachophoresis of Immunoglobulins
• Chapter 8: The Chemical Modification of Proteins, Haptens, and Solid Supports
• Chapter 9: Reagents for Immunofluorescence and Their Use for Studying Lymphoid Cell Products
• Chapter 10: Radiolabeling and Immunoprecipitation of Cell-Surface Macromolecules
• Chapter 11: Haptenation of Viable Biological Carriers
• Chapter 12: Production and Assay of Murine Anti-Allotype Antisera
• Chapter 13: Preparation of Mouse Antisera against Histocompatibility Antigens
• Chapter 14: Technique of HLA Typing by Complement-Dependent Lympholysis
• Chapter 15: The MLR Test in the Mouse
• Chapter 16: A Sensitive Method for the Separation of Rosette-Forming Cells
• Chapter 17: The Use of Protein A Rosettes to Detect Cell-Surface Antigens
• Chapter 18: Hapten–Gelatin Gels Used as Adsorbents for Separation of Hapten-Specific B Lymphocytes
• Chapter 19: Assay for Plaque-Forming Cells
• Chapter 20: Plaquing and Recovery of Individual Antibody-Producing Cells
• Chapter 21: Assay for Specific Alloantigen-Binding T Cells Activated in the Mixed Lymphocyte Reaction
• Chapter 22: Assay for Antigen-Specific T-Cell Proliferation in Mice
• Chapter 23: Antigen-Specific Helper T-Cell Factor and Its Acceptor
• Chapter 24: In Vitro Immunization of Dissociated Murine Spleen Cells
• Chapter 25: Induction of a Secondary Antibody Response in Vitro with Rabbit Peripheral Blood Lymphocytes
• Chapter 26: Induction of Immune Responses with Clonal Dominance at High Antibody Levels
• Chapter 27: Limiting Dilution Analysis
• Chapter 28: Establishment and Maintenance of Murine Lymphoid Cell Lines in Culture
• Chapter 29: Clonal Growth of Cells in Semisolid or Viscous Medium
• Chapter 30: Preparation of Sendai Virus for Cell Fusion
• Chapter 31: Fusion of Lymphocytes
• Chapter 32: Soft Agar Cloning of Lymphoid Tumor Lines: Detection of Hybrid Clones with Anti-SRBC Activity
• Chapter 33: Isotope Laboratory
• Chapter 34: Analysis of Immunological Data
• Subject Index