Contents

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

DavidHughes 173

14. Immunoelectron Microscope Techniques in Plant Virus Diagnosis DaphneM. 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

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