Immunological Methods - 1st Edition - ISBN: 9780124427037, 9781483269634

Immunological Methods

1st Edition

Volume III

Editors: Ivan Lefkovits Benvenuto Pernis
eBook ISBN: 9781483269634
Imprint: Academic Press
Published Date: 8th October 1985
Page Count: 506
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Immunological Methods, Volume III provides information pertinent to the fundamental aspects of immunological methods. This book presents recombinant DNA technology as applied to immunology.

Organized into 25 chapters, this volume begins with an overview of the major histocompatibility complex. This text then examines the use of monoclonal antibodies in the identification of lymphocyte membrane antigens and explains the use of monoclonal antibodies in histocompatibility typing. Other chapters consider the methods in two-dimensional electrophoretic analysis of proteins and another to lymphokines that support B cells. This book discusses as well the production and expansion of cloned B and T cell lines and hybridomas. The final chapter deals with the sophisticated methodology used with particular animal species, namely, birds, sheep, and amphibians to probe specific immunological questions.

This book is a valuable resource for immunologists and investigators with extensive experience in using immunological methods. Protein chemists and biologists will also find this book useful.

Table of Contents



Contents of Previous Volumes

Abbreviations List

1 Methods in Molecular Immunology: Chromosomal Walking in the Major Histocompatibility Complex

I. Introduction

II. Materials

III. Construction of Cosmid Libraries

IV. Screening of Cosmid Libraries

V. Characterization of Cosmid Clones

VI. Appraisal


2 DNA-Mediated Transformation of Lymphoid Cells

I. Introduction

II. Dominant Selectable Markers

III. Techniques of Stable Gene Transfer

IV. Other Techniques of Stable Gene Transfer


3 Cloning of cDNAs with Vectors Permitting Expression in Host Cells

I. Introduction

II. General Strategies and Their Problems

III. Experimental Procedures

IV. Some Comments


4 Construction of Vectors for Immunoglobulin Reverse Genetics

I. Introduction

II. General Strategy

III. Material and Stock Solutions

IV. Experimental Procedures

V. Examples of Constructions


5 Strategies of Oligonucleotide Synthesis

I. Introduction

II. Strategies

III. Experimental Section

IV. Reagents and Solvents That Must Be Purified or Prepared


6 High-Performance Liquid Chromatography of Biologically Active Proteins in the Nanogram (Picomole) Range

I. Introduction

II. Materials and Methods

III. Separation by Size: Gel Permeation

IV. Separation by Charge: Ion Exchange and Hydroxyapatite

V. Separation by Hydrophobicity: Reverse Phase

VI. Quantitation and Limits of Detection

VII. Protein Purification


7 The HPLC of Immunoglobulins

I. Introduction

II. HPLC: Theory, Practice, and Equipment

III. Immunoglobulins on HPLC

IV. Discussion


8 Preparation of Liposomes Carrying Lymphocyte Membrane Proteins

I. Introduction

II. Isolation of Membrane Proteins

III. Preparation of Liposomes

IV. Applications


9 Detection of Glycolipid Antigens with Monoclonal Antibodies

I. Introduction

II. Preliminary Tests

III. Lipid Extraction

IV. Solid-Phase RIA

V. Hapten Inhibition and Hapten Binding

VI. Thin-Layer Chromatography

VII. Critical Appraisal


10 Mapping of Novel B Lymphocyte Differentiation Antigens

I. Introduction

II. Definition of B Cell Differentiation Antigens by Negative Selection Assay

III. Two-Dimensional Gel Analysis of B Cell Differentiation Antigens

IV. Appendix


11 Use of Large-Scale Two-Dimensional ISODALT Gel Electrophoresis System in Immunology

I. Objective

II. Materials

III. Methods

IV. Critical Appraisal


12 Tissue Typing Using Biosynthetically Labeled Monoclonal Antibodies

I. Introduction

II. Biosynthetic Labeling

III. Purification

IV. Activity and Stability

V. Binding Assay

VI. Applications and Limitations


13 Assays for Lymphokines Supporting B Cell Growth

I. Introduction

II. Materials

III. Preactivation Conditions

IV. Recultivation Conditions

V. Measurement and Proliferation


14 Establishment of Cellular Microenvironments for in Vitro B Cell Progenitor Differentiation

I. Introduction

II. Principle of the Method

III. Materials

IV. Procedure

V. Comments


15 Development of B Cell Progenitors in Semisolid Agar Cultures

I. Introduction

II. Experimental Methods

III. Summary of Results


16 Isolation of Human Alloreactive T Helper Clones and Their Use as Polyclonal Activators of B Cells

I. Objective and Principle of the Method

II. Materials

III. Procedure

IV. Critical Appraisal


17 Cytolytic T Cell Clones and Hybridomas

I. Introduction

II. Materials

III. Methods

IV. Application


18 Conditions for the Establishment in Vitro of Interleukin 3-Dependent Murine B Cell Precursor Lines

I. Introduction

II. Materials

III. WEHI-3 Supernatants

IV. Assay for IL-3 Activity

V. Preparation of Cells

VI. Establishment of IL-3-Dependent B Cell Precursor Lines

VII. Cloning IL-3-Dependent B Cell Precursors

VIII. Characterizing the Cell Lines

IX. Concluding Remarks


19 Assays for the Detection of Rheumatoid Factor-Producing Cells

I. Introduction

II. Hemolysis Assay

III. Plaque Assay

IV. Radioimmunoassay


20 Fluorescence Cell Sorter Techniques in Immunology

I. Introduction

II. Description of a Cell Sorter

III. Preparation of Reagents

IV. Sample Preparation

V. Cell Separation

VI. Data Acquisition

VII. Single-Parameter Data Analysis and Interpretation

VIII. Examples

21 Tables for Evaluating Limiting Dilution Experiments

I. Introduction

II. Percentage of Single Clones

III. Confidence Limits

IV. Test for Independence in 2 x 2 Contingency Tables


22 Development of an Avian Lymphoid Chimera

I. Introduction

II. Principle of the Method

III. Details of the Procedure

IV. Evaluation of Chimeras

V. Comments


23 Avian Embryos in Immunology

I. Introduction

II. Materials

III. Methods


24 Sheep as an Experimental Model for Immunology: Immunological Techniques in Vitro and In Vivo

I. Introduction

II. Maintenance of Experimental Sheep

III. In Vivo Procedures

IV. In Vitro Procedures


25 Methods Used to Study the Immune System of Xenopus (Amphibia, Anura)

I. Introduction: The Immune System of Xenopus

II. Strains and Species Available

III. Operating on Xenopus

IV. Preparing Lymphocytes for Cell Culture

V. Making Reagents

VI. Analyzing the Immune System

VII. Functional Assays Analyzing Immune Response

VIII. Somatic Cell Hybrid Between Frog Lymphocytes and Mouse Myeloma Cells

IX. Cell Markers, Ploidy Markers

X. Appendix




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© Academic Press 1985
Academic Press
eBook ISBN:

About the Editor

Ivan Lefkovits

Affiliations and Expertise

Basel Institute for Immunology, Basel, Switzerland

Benvenuto Pernis

Ratings and Reviews