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A2926

Antibodies: A Laboratory Manual

extensive notes and two-color illustrations

  •  ISBN-10 0-87969-314-2

  •  ISBN-13 978-0-87969-314-5

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Properties

publication info   E. Harlow and D. Lane, Cold Spring Harbor Laboratory Press, 1988, 726 pp., comb bound
mfr. no.   Cold Spring Harbor

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.

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

Safety & Documentation

Safety Information

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Documents

Certificate of Analysis (COA)

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Protocols & Articles

Protocols

ELISA Procedures

To search our entire portfolio of antibodies for your research needs, use our Antibody Explorer Tool.
Keywords: Capture ELISA, Cell culture, Cell signaling, Enzyme-linked immunosorbent assay, Hormones, Indirect ELISA, Titrations

Immunofluorescence Labeling of Cells

To search our entire portfolio of antibodies for your research needs, use our Antibody Explorer Tool.
Keywords: Microscopy, Solvents, Titrations

Immunoprecipitation Procedure

To search our entire portfolio of antibodies for your research needs, use our Antibody Explorer Tool.
Keywords: Buffers, Cell disruption, Centrifugation, Enzyme activity, Immobilization, Immunoprecipitation, Individual protein Immunoprecipitation, PAGE, Size-exclusion chromatography, Western blot

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