Immunochemical protocols

Preface. .............................................................................................................v Contributors .....................................................................................................ix 1. Immunization Strategies for Antibody Production Robert Burns ......................................................................................... 1 2. Synthetic Peptides as Antigens for Antibody Production David C. Hancock and Nicola J. O’Reilly .......................................... 13 3. Production of Polyclonal Antibodies in Rabbits David C. Hancock and Nicola J. O’Reilly .......................................... 27 4. Making Hybridomas Robert Burns ....................................................................................... 41 5. Growing Hybridomas Gary Entrican and Gareth Young ....................................................... 55 6. Phage Display Vectors for the In Vitro Generation of Human Antibody Fragments Michael Hust and Stefan Dübel .......................................................... 71 7. Purification of Antibodies and Preparation of Antibody Fragments Graham D. Kemp ................................................................................ 97 8. Conjugation of Antibodies to Alkaline Phosphatase G. Brian Wisdom .............................................................................. 123 9. Conjugation of Antibodies to Horseradish Peroxidase G. Brian Wisdom .............................................................................. 127 10. Conjugation of Antibodies to Fluorescein or Rhodamine G. Brian Wisdom .............................................................................. 131 11. Conjugation of Antibodies to Biotin James D. Hirsch and Rosaria P. Haugland ....................................... 135 12. Preparation of Colloidal Gold Probes David Hughes.................................................................................... 155 13. Immunogold Probes in Light Microscopy David Hughes.................................................................................... 173 14. Immunoelectron Microscope Techniques in Plant Virus Diagnosis Daphne M. Wright ............................................................................ 193 15. Dual-Antibody Sandwich Enzyme-Linked Immunosorbent Assay William Jordan .................................................................................. 207 16. Competitive Enzyme-Linked Immunosorbent Assay William Jordan .................................................................................. 215 17. Immunoblotting Techniques Barbara Magi and Sabrina Liberatori ............................................... 227 18. Epitope Mapping Glenn E. Morris ................................................................................. 255 19. Generation and Testing of Immunoaffinity Columns Weilin L. Shelver ............................................................................... 269 20. Immunocapture-Polymerase Chain Reaction Vincent Mulholland .......................................................................... 281 21. Magnetic Cell Sorting Maciej Zborowski and Jeffrey J. Chalmers ....................................... 291 22. Cell Enrichment and Immunochemical Staining Oscar R. Lara and Jeffrey J. Chalmers .............................................. 301 Index...............................................................................................................311