Immunochemical Protocols

ISBN-13: 9781617374661 / Angielski / Miękka / 2010 / 318 str.

Robert Burns

Opis

Immunochemical techniques have been in use for many years with early examples of bacterial strain typing dating back to the 1940s. The basis for the science is the exquisite elegance of the mammalian immune system with its ability to recognize foreign proteins and to manufacture antibody m- ecules that strongly bind to the substances that elicited them. Not only are potentially harmful pathogens and toxins recognized by the immune system, but the system can be persuaded to manufacture antibodies to an astonishing array of substances. In the early days of this science, all antibodies for investigative work were produced by immunizing mammals with the substance of interest, followed by regular donor bleeds that yielded antisera. Serum produced in this way yields heterogenic populations of antibody molecules recognizing different epitopes on the target protein, which may be adequate for its intended p- poses, but can also cause problems of crossreactivity. In 1975, Kohler and Milstein reported that spleen cells from immune donor animals could be immortalized, cloned from single cells, and grown in continuous culture. This original work described the method for the production of monoclonal antibodies.

Recenzje

Reviews of Previous editions:
"...a treasury of detailed information, both within the protocols and in the Notes, which in some chapters are extensive...will be useful to both those who already have some experience in this area and...to those who wish to embark on new techniques."
-Microbiology Today

An excellent and well arranged compilation of a range of antibody techniques....An excellent laboratory manual.-Immunoassay

Spis treści

Immunization Strategies for Antibody Production Robert Burns Synthetic Peptides as Antigens for Antibody Production David C. Hancock and Nicola J. O'Reilly Production of Polyclonal Antibodies in Rabbits David C. Hancock and Nicola J. O'Reilly Making Hybridomas Robert Burns Growing Hybridomas Gary Entrican and Gareth Young Phage Display Vectors for the In Vitro Generation of Human Antibody Fragments Michael Hust and Stefan Dübel Purification of Antibodies and Preparation of Antibody Fragments Graham D. Kemp Conjugation of Antibodies to Alkaline Phosphatase G. Brian Wisdom Conjugation of Antibodies to Horseradish Peroxidase G. Brian Wisdom Conjugation of Antibodies to Fluorescein or Rhodamine G. Brian Wisdom Conjugation of Antibodies to Biotin James D. Hirsch and Rosaria P. Haugland Preparation of Colloidal Gold Probes David Hughes Immunogold Probes in Light Microscopy David Hughes Immunoelectron Microscope Techniques in Plant Virus Diagnosis Daphne M. Wright Dual-Antibody Sandwich Enzyme-Linked Immunosorbent Assay William Jordan Competitive Enzyme-Linked Immunosorbent Assay William Jordan Immunoblotting Techniques Barbara Magi and Sabrina Liberatori Epitope Mapping Glenn E. Morris Generation and Testing of Immunoaffinity Columns Weilin L. Shelver Immunocapture-Polymerase Chain Reaction Vincent Mulholland Magnetic Cell Sorting Maciej Zborowski and Jeffrey J. Chalmers Cell Enrichment and Immunochemical Staining Oscar R. Lara and Jeffrey J. Chalmers Index

Blurb

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.