Burns Immunochemical Protocols

3rd Auflage 2005
ISBN: 978-1-59259-873-1
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 295, 318 Seiten, eBook

Reihe: Methods in Molecular Biology

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.

Zielgruppe

Research

Autoren/Hrsg.

Burns, Robert

Inhaltsverzeichnis

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.