A2926
Sigma
Antibodies: A Laboratory Manual
| ISBN-10: | 0-87969-314-2 |
| ISBN-13: | 978-0-87969-314-5 |
Description
| General description | This book places the production and use of polyclonal and monoclonal antibodies within the reach of every biologist. All of the techniques described are supplemented with extensive notes and two-color illustrations. |
Properties
| publication info | E. Harlow and D. Lane, Cold Spring Harbor Laboratory Press, 1988, 726 pp., comb bound |
Table Of Contents
| Table of Contents | Immune Response, An introduction to immunologic principles Antibody Molecules–review of antibody structure and genetics of immunoglobulin genes Antibody-Antigen Reactions–review of structure of interaction, affinity, avidity Antibody Response–how it works; Immunizations–immunogenicity, preparing the antigen, immunization protocols, collecting sera, how to make weak antigens strong Monoclonal Antibodies–step-by-step production–immunization, developing screening methods, production of hybridomas Growing Hybridomas–growing and maintaining myeloma and hybridoma cell lines, producing and collecting monoclonal antibodies, methods for drug selection Storing and Purifying Antibodies Labeling Antibodies Cell Staining–constraints, choice of antibody, protocols for staining Immunoprecipitation–constraints, choice of antibody, protocols Immunoaffinity Purification–constraints, choice of antibody, protocols Immunoassay–types of immunoassays, where to start, antibody capture assays, antibody sandwich assays, antigen capture assays, detection, designing immunoassays Reagents–bacterial cell wall proteins that bind antibodies, anti-immunoglobulin antibodies, proteolytic fragments of antibodies, acetone powders Appendices Electrophoresis¿one- and two-dimensional gels, alternate buffer systems, silver and Coomassie staining techniques, fluorography, radiography; Protein Techniques–tables, quantitation methods, TCA precipitation; General Information–buffers, stock solutions, detergents; Bacterial Expression–techniques for screening, vectors for bacterial expression Features Summary Tables–help you decide strategy for experimental course most suited to your objective; Constraints Indicated–what can and cannot work Help in Choosing Proper Antibodies for Each Technique Illustrative Keys–aid in finding proper sequence of techniques Extensive Advice Based on Authors' Experience Trouble-shooting–help in avoiding experimental pitfalls and discussion of likely problems and their solution Cautions–safety notes throughout protocols Techniques Detailed Protocols for the Production of Polyclonal and Monoclonal Antibodies–tissue culture supernatants and ascites Purification of Antibodies–purification of polyclonal and monoclonal antibodies using a broad range of conventional techniques, protein A-based techniques, affinity chromatography Techniques for Labeling Purified Antibodies with Iodine, Enzymes, Fluorochromes, and Biotin–relative advantages and disadvantages for each label Cell Staining for Tissue Culture Cells, Yeast, and Tissue Sections from the Preparation of Samples Through Detection Immunoprecipitation–labeling, lysis, and purification of immune complexes with extensive trouble-shooting section Immunoblotting–from preparation of samples through detection Immunoaffinity Purification–multiple examples of techniques for preparation of antibody columns and suggested strategies |
