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The now classic lab manual Antibodies, by Harlow and Lane, has been revised, extended, and updated by Edward Greenfield of the Dana-Farber Cancer Center, with contributions from other leaders in the field. This second edition of the manual is an essential resource for molecular biology, immunology, and cell culture labs on all matters relating to antibodies. The chapters on hybridomas and monoclonal antibodies have been recast with extensive new information and there are additional chapters on characterizing antibodies, antibody engineering, and flow cytometry. As in the original book, the emphasis in this second edition is on providing clear and authoritative protocols with sufficient background information and troubleshooting advice for the novice as well as the experienced investigator.

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Preface

1 Antibody Production by the Immune System

Stefanie Sarantopoulos

2 The Antibody Molecule

Stefanie Sarantopoulos

3 Antibody-Antigen Interactions

Stefanie Sarantopoulos

4 Antibody Responses

Stefanie Sarantopoulos

5 Selecting the Antigen

Edward A. Greenfield, James DeCaprio, and Mohan Brahmandam

6 Immunizing Animals

Edward A. Greenfield

7 Generating Monoclonal Antibodies

Edward A. Greenfield

8 Growing Hybridomas

Edward A. Greenfield

9 Characterizing Antibodies

Frances Weis-Garcia and Robert H. Carnahan

10 Antibody Purification and Storage

Jordan B. Fishman and Eric A. Berg

11 Engineering Antibodies

James Dasch and Amy Dasch

12 Labeling Antibodies

Eric A. Berg and Jordan B. Fishman

13 Immunoblotting

Larisa Litovchick

14 Immunoprecipiation

James DeCaprio and Thomas O. Kohl

15 Immunoassays

Thomas O. Kohl and Carl A. Ascoli

16 Cell Staining

Scott J. Rodig

17 Antibody Screening using High Throughput Flow Cytometry

Thomas D.l. Duensing and Susan R. Watson

Appendix I: Electrophoresis

Appendix II: Protein Techniques

Appendix III: General Information

Appendix IV: Bacterial Expression

Appendix V: Cautions

Index