Anmerkung: Front Cover -- Methods in Cell Biology, Volume 37 -- Copyright Page -- Contents -- Contributors -- Preface -- Chapter 1. The Immune Response: An Overview -- I. Clonal Selection -- II. Size of the Immune Repertoire -- III. Specificity and Affinity Maturation -- IV. Idea of Cross-Reactivity -- V. Monoclonality and Multispecificity -- VI. A Cautionary Note -- Suggested Readings -- Chapter 2. Production of Polyclonal Antibodies against Proteins and Peptides -- I. Introduction -- II. Properties of Immunoglobulins -- III. Polyclonal versus Monoclonal Antibodies -- IV. Processing of Antigens for Immunization -- V. Adjuvants -- VI. Routes of Immunization -- VII. General Schedule for Immunization -- VIII. Collection of Blood -- IX. Preparation, Storage, and Shipping of Serum -- X. Determination of Antibody Titer (Test of Reactivity) -- XI. Test of Specificity -- XII. Production and Selection of Isofom-Specific Immunoglobulins -- XIII. Purification of Immunoglobulins -- References -- Chapter 3. Making Monoclonal Antibodies -- I. lntroduction -- II. First Principles -- III. Overview of Protocol -- IV. Hybridoma Protocols -- V. Perspective -- References -- Chapter 4. Purification of Mouse Antibodies and Fab Fragments -- I. Introduction -- II. Purification of Murine IgG Monoclonal Antibodies -- III. Generation of Fab and Fab' Fragments -- IV. Fab Purification -- V. Further Purification of Fab Fragments -- VI. Crystallization of Fab Fragments -- VII. Summary -- References -- Chapter 5. Blot-Affinity Purification of Antibodies -- I. Introduction -- II. SDS-Polyacrylamide Gel Electrophoresis and Blotting -- Ill. Antibody Adsorption, Elution, and Storage -- IV. Quality Evaluation of the Eluted Antibodies -- V. Discussion -- References -- Chapter 6. Immunoblotting -- I. Introduction -- II. Semidry and Wet Blotting of Gels. , III. Staining Nitrocellulose with Antibodies -- IV. Miscellaneous Information -- V. Perspectives -- References -- Chapter 7. Immunoprecipitation Methods -- I. Introduction -- II. Reagents and Solutions -- III. Protocol for Immunoprecipitation -- IV. Troubleshooting -- References -- Chapter 8. A Quantitative Solid-Phase Binding Assay for Tubulin -- I . Introduction -- II. Materials -- III. Methods -- IV. Typical Results and Discussion -- V. Development of a Competitive ELISA -- References -- Chapter 9. Confocal Immunofluorescence Microscopy of Microtubules in Oocytes, Eggs, and Embryos of Algae and Amphibians -- I . Introduction -- II . Immunofluorescence Microscopy of Pelvetia and Xenopus -- III. Confocal Microscopy: Advantages and Considerations -- IV. Conclusion -- Appendix I: Recipes and Reagents -- Appendix II: Antibodies -- Appendix III: Supplier Information -- References -- Chapter 10. Immunolabeling of Antigens in Plant Cells -- I. Introduction -- II. Reagents -- III. Immunofluorescence Microscopy of Cytoskeletal Elements in Isolated Root Tip Cells -- IV. Preparation of Leaf Epidemial Sheets for Immunolabeling -- V. Cryosectioning for Light Microscopy Immunolabeling -- VI. Troubleshooting/Variations on Protocols -- VII. Perspectives -- References -- Chapter 11. Antigen Localization in Fission Yeast -- I . Introduction -- II. Immunofluorescence Microscopy -- III. Solutions and Notes for Immunofluorescence Microscopy -- IV. Fluorescence Microscopy -- V. Solutions and Notes for Fluorescence Microscopy -- VI. Immunogold Labeling of Yeast Ultrathin Sections -- References -- Chapter 12. Nonfluorescent Immunolocalization of Antigens in Mitotic Sea Urchin Blastomeres -- I. Introduction -- II. Immunolocalization and Sea Urchin Embryos -- III. Reagents and Buffers -- IV. Antibody Preparation -- V. Preparation of Sea Urchin Embryos -- VI. Immunostaining. , VII. Alternative Techniques -- VIII. Kesults and Perspectives -- References -- Chapter 13. Ultrasmall Gold Probes: Characteristics and Use in Immuno(cyto)chemical Studies -- I. Introduction -- II. Improvement of Detection System -- III. Characteristics of Ultrasmall Gold Probes -- IV. Application of Ultrasmall Gold Probes -- V. Discussion -- References -- Chapter 14. Immunogold Electron Microscopy: Mapping Tubulin Isotypes on Neurite Microtubules -- I. Introduction -- II. Scope and Application -- III. Labeling Protocols (Preembedding) -- IV. Results and Perspectives -- V. Controls and Critical Considerations -- VI. Troubleshooting Guide -- VII. Conclusion -- VIII. Appendix -- References -- Chapter 15. Postembedding Immunocytochemical Techniques for Light and Electron Microscopy -- I. Introduction -- II. Protocols -- III. Recipes -- References -- Chapter 16. Electron Microscopy Immunocytochemistry Following Cryofixation and Freeze Substitution -- I. Introduction -- II. Systems and Goals -- III. Cryofixation, Freeze Substitution, and Embedding -- IV. Antibodies and Gold Markers -- V. Protocols -- VI. Conclusions and Perspectives -- References -- Chapter 17. Microinjection of Antibodies in the Analysis of Cellular Architecture and Motility -- I. Introduction: The Scope of the Play -- II. Actors and Requisites in the Play -- III. The Set -- IV. The Performance -- V. The Lessons to Be Learned -- VI. Perspectives -- References -- Chapter 18. Immunoselection and Characterization of cDNA Clones -- I . Introduction -- II. Overview of a Typical Immunoselection Project -- III. Preparation of Antibodies for Library Screening -- IV. Choice and Preparation of Expression Libraries -- V. Immunoselection of Clones from cDNA Librariesrs -- VI. Characterization of Cloned cDNAs -- VII. Cloned cDNAs as Hybridization Probes -- VIII. Fusion Protein Analysis. , IX. Confirmation of cDNA Clones -- X. Sequence Analysis of Cloned cDNAs -- XI. Materials and Reagents -- References -- Chapter 19. Antiidiotypic Antibodies: Methods, Applications, and Critique -- I . Introduction -- II. Structural Basis and Main Principles of Antiidiotypic Approach -- III. Production and Characterization of Antiidiotypic Antibodies -- IV. Receptor Identification by Antiidiotypic Antibodies -- V. Applications: Use of Antiidiotypic Antibodies to Study Molecular Interactions of Intermediate Filament Proteins -- VI. Conclusions -- References -- Appendix -- Index -- Volumes in Series.