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163,40 €Preface
1 Antibody Production by the Immune System
2 The Antibody Molecule
3 Antibody-Antigen Interactions
4 Antibody Responses
5 Selecting the Antigen
6 Immunizing Animals
7 Generating Monoclonal Antibodies
8 Growing Hybridomas
9 Characterizing Antibodies
10 Antibody Purification and Storage
11 Engineering Antibodies
12 Labeling Antibodies
13 Immunoblotting
14 Immunoprecipiation
15 Immunoassays
16 Cell Staining
17 Antibody Screening using High Throughput Flow Cytometry
Appendix I: Electrophoresis
Appendix II: Protein Techniques
Appendix III: General Information
Appendix IV: Bacterial Expression
Appendix V: Cautions
Index
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
As Dr. Greenfield notes in his preface to this second edition: "The Antibodies manual provided our laboratory with guidance in the form of protocols and recommendations for setting up a hybridoma facility. Everything we needed to know to make a monoclonal antibody was all there, neatly packaged in an easy-to-understand book....the second edition is intended to provide the necessary information and protocols to assist investigators with their first monoclonal antibody effort as well as to provide guidance for more experiences antibody makers who are having some difficulties with a particular project."
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