Laboratory techniques in biochemistry and molecular biology. Volume 13, Monoclonal antibody technology: the production and characterization of rodent and human hybridomas /

This volume contains detailed, comprehensive advice on rat, mouse and human hybridoma production. It begins with a general introduction, then describes the practical applications of the technology with photographs and protocols for everything from animal dissection to epitope analysis of antigens.

Detalles Bibliográficos
Clasificación:Libro Electrónico
Autor principal: Campbell, Ailsa M.
Formato: Electrónico eBook
Idioma:Inglés
Publicado: Amsterdam ; Oxford : North-Holland Pub. Co., 1984.
Colección:Laboratory techniques in biochemistry and molecular biology ; 13.
Temas:
Biochemistry > Technique.
Molecular biology > Technique.
Biochimie > Technique.
Biologie mol�eculaire > Technique.
Acceso en línea:Texto completo
Tabla de Contenidos:
  • Front Cover; Monoclonal Antibody Technology; Copyright Page; Contents; Preface; Acknowledgements; Abbreviations; Chapter 1. General properties and applications of monoclonal antibodies; 1.1. Introduction; 1.2. Comparison of monoclonal antibodies and conventional antiserum; 1.3. Application of monoclonal antibodies; Chapter 2. Assay techniques; 2.1. General assay requirements; 2.2. Theoretical considerations; 2.3. Practical considerations; 2.4. Antibody sampling; 2.5. Types of assay; 2.6. Solid-phase-assays; 2.7. Soluble-phase systems; 2.8. Cellular assays; 2.9. Biological assays
  • 2.10 Immunocytochemical assaysChapter 3. Selection of animals and cell lines; 3.1. Choice of animal; 3.2. Choice of cell lines; 3.3. Selective drug markers; 3.4. The mouse system; 3.5. The rat system; 3.6. The human system; 3.7. T cell lines; Chapter 4. Immunisation; 4.1. Introduction; 4.2. Immunisation schedules; 4.3. Immunisation in vitro; 4.4. Isotype enrichment; Chapter 5. Cell culture requirements for hybridomas; 5.1. Introduction; 5.2. Basic requirements; 5.3. Basic cell-culture techniques; 5.4. Contamination; 5.5. Feeder cells; Chapter 6. Fusion procedures
  • 6.1. The use of polyethylene glycol6.2. The components of HAT medium; 6.3. Selection of hybridomas by fluorescence activated cell sorting; 6.4. Preparation of stock solutions; 6.5. Fusion frequencies and plating densities; 6.6. Fusion protocols for mouse experiments; Chupter 7. Transformation; 7.1. Introduction; 7.2. Pre-selection of lymphocytes of pre-defined specificity; 7.3. Pre-selection of T lymphocytes; 7.4. Transformation; 7.5. Transformation followed by fusion; Chapter 8. Selection and cloning; 8.1. Early feeding and assay of fusions; 8.2. Failure of fusions
  • 8.3. Cloning of hybridomas8.4. Failure of cloning; 8.5. Continuation of cloning; Chapter 9. Antibody production and purification; 9.1. Maintenance of cell stocks; 9.2. Expansion of hybridomas in vitro; 9.3. Expansion of hybridomas in vivo; 9.4. Failure in expansion; 9.5. Storage of antibody; 9.6. Concentrating the antibody; 9.7. Purification of the antibody; Chapter 10. Characterisation of monoclonal antibodies; 10.1. Introduction; 10.2. Determination of antibody class; 10.3. Proof that the antibody is monoclonal; 10.4. Epitope analysis; 10.5. Determination of antibody affinity
  • 10.6. Karyotype analysis of hybridomasAppendix 1. Animal handling techniques; Appendix 2. Addresses of suppliers and manufacturers; Appendix 3. Protocols for polyacrylamide gel electrophoresis (PAGE); References; Addendum; Subject Index

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