© 2014 • 847 pp., illus. (32 4C, 103 B&W), index
Paperback • $147.00 • ISBN 978-1-936113-81-1

It has been 25 years since the now classic lab manual Antibodies, by Harlow and Lane appeared. This second edition of Antibodies, edited by Edward A. Greenfield of the Dana-Farber Cancer Institute builds on the core strengths of the first edition, presenting clear and authoritative protocols with extensive background information and troubleshooting advice. The original introductory chapters have been recast and updated to take into account our current understanding of the immune system. The critical chapters on generating monoclonal antibodies and growing hybridomas, which demystified hybridoma generation, have been greatly expanded and updated to make these procedures easy to follow and adaptable to current research needs. The remaining chapters contain all new protocols and topics and reflect the progress in how antibodies are studied and used since the appearance of the original edition. These include:

• antibody purification and storage
• engineering antibodies, including use of degenerate oligonucleotides, 5’-RACE, phage display, and mutagenesis
• extensive labeling techniques
• new immunoblotting protocols
• the latest screening and labeling techniques

What’s New in This Edition?

I have the first edition. Why should I buy the second edition?

While the core of the first edition appears in updated and recast form in the second edition, more than half of the second edition contains entirely new information and protocols to reflect the changes in the field since the first edition.

To whom is this edition addressed? Is this a book for immunologists?

As with the first edition, the second edition provides information and protocols for both the neophyte and the experienced investigator. It dissects the mysteries of producing the antibodies and reagents you need for your experiments with the needs of molecular biologists in mind. It is the perfect book for new members of your lab as it has sufficient background information to provide context as well as clear, step-by-step instructions for performing the needed experiments—both the why and the how for these techniques. And if things go wrong, there is extensive troubleshooting to diagnose the problem.

Are the protocols in this book better than those available for download on the Web?

Many of the protocols available on the Web are perfectly good, but there are many that are not. Reagents are expensive and deadlines are tight. CSHL Press manuals provide protocols that have been formulated and tested in the labs of leading investigators in the field. They are reliable and they work. And they include the context and troubleshooting information that many online protocols do not have.

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