Immunochemical Protocols

Guidelines and Methods for Successful
Antibody Design and Production

This new edition of a much praised classic includes both basic method for producing and using antibodies and advanced techniques required by today's state-of-the art investigators. The basic sections of the book stress the first principles of antibody production and guide the reader through the processes and pitfalls of immunochemistry. The advanced sections focus on specific applications and cover the immunochemical techniques now used in laboratories. New material includes techniques for making new monoclonal cell lines, for producing good quality monoclonal antibodies, immunocapture PCR, immunoaffinity purification, and immunoelectron microscopy in plant virus diagnosis.

Includes supplementary material: sn.pub/extras

Klappentext
This new third edition of a much praised classic has been updated and revised to provide both basic methods for producing and using antibodies and advanced techniques required by today's state-of-the art investigators. The basic sections of the book stress the first principles of antibody production and guide the reader through the processes and pitfalls of immunochemistry. The advanced sections focus on specific applications and cover the major immunochemical techniques currently used in laboratories. New material includes techniques for making new monoclonal cell lines, for producing good quality monoclonal antibodies, immunocapture PCR, immunoaffinity purification, and immunoelectron microscopy in plant virus diagnosis. The authors provide full details of methods for antigen selection and preparation, antibody production, and reagent manufacture. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Up-to-date and highly practical, Immunochemical Protocols, Third Edition, takes both novice and experienced researchers from project initiation to final test format with a broad array of protocols suited to immediate use in the laboratory, as well as methods that can be successfully applied to entirely new systems and processes.

Inhalt
Immunization Strategies for Antibody Production.- Synthetic Peptides as Antigens for Antibody Production.- Production of Polyclonal Antibodies in Rabbits.- Making Hybridomas.- Growing Hybridomas.- Phage Display Vectors for the In Vitro Generation of Human Antibody Fragments.- Purification of Antibodies and Preparation of Antibody Fragments.- Conjugation of Antibodies to Alkaline Phosphatase.- Conjugation of Antibodies to Horseradish Peroxidase.- Conjugation of Antibodies to Fluorescein or Rhodamine.- Conjugation of Antibodies to Biotin.- Preparation of Colloidal Gold Probes.- Immunogold Probes in Light Microscopy.- Immunoelectron Microscope Techniques in Plant Virus Diagnosis.- Dual-Antibody Sandwich Enzyme-Linked Immunosorbent Assay.- Competitive Enzyme-Linked Immunosorbent Assay.- Immunoblotting Techniques.- Epitope Mapping.- Generation and Testing of Immunoaffinity Columns.- Immunocapture-Polymerase Chain Reaction.- Magnetic Cell Sorting.- Cell Enrichment and Immunochemical Staining.