ELISA: Theory and Practice: Methods in Molecular Biology, Vol. 42

Features and Benefits

The book provides readers with full descriptions of the basic systems that make ELISA one of the most powerful techniques in science today. The editor′s goal is to provide enough information for a full appreciation of theory, and enough practical exercises for a thorough facility with lab operation.

Table of Contents

Basic Immunology: 1. The Immune Response. 2. Antigens. 3. Adaptive Immunity and Clonal Selection. 4. Antibodies. 5. Antigenic Commonness.
Basic Principles of ELISA: 1. Reaction Schemes. 2. Direct ELISA. 3. Indirect ELISA. 4. Sandwich ELISA. 5. Competition ELISA. 6. Choice of Assays.
Stages in ELISA: 1. Solid-Phase. 2. Washing. 3. Addition of Reagents. 4. Incubation. 5. Blocking Conditions and Nonspecific Reactions. 6. Enzyme Conjugates. 7. Availability of Conjugates. 8. Conjugation with Enzymes. 9. Development of Label. 10. Stopping Reactions. 11. Reading. 12. Practical Problems.
Theoretical Considerations: 1. Setting Up and Use of ELISA. 2. What Is Already Known. 3. Complexity of Problems. 4. Antigenic Considerations. 5. Other Techniques. 6. Units. 7. Making Dilutions. 8. Pipettes. 9. Molarities.
Direct ELISA: Titration of Antigen and Antibody: 1. Learning Principles. 2. Reaction Scheme. 3. Basis of Assay. 4. Materials and Equipment. 5. Practical Details. 6. Data Explained. 7. Aspects of Assay Described. 8. Conclusions.
Indirect ELISA: 1. Learning Principles. 2. Basis of Assay. 3. Materials and Reagents. 4. Practical. 5. Use of Indirect ELISA to Titrate Antibodies. 6. Use of Indirect ELISA to Determine the Positivity of Sera at Single Dilution.
Use of Antibodies on Solid-Phase in Capture ELISA: 1. Use of Capture ELISA to Detect and Titrate Antigen. 2. Reaction Scheme. 3. Basis of Assay. 4. Materials. 5. Methods. 6. Titration of Capture Antibody Using IgG. 7. Titration of Capture Antibody when Used as Whole Serum. 8. Use of Capture ELISA to Detect Antigens. 9. Use of Capture ELISA to Detect and Titrate Antibodies.
Competitive ELISA: 1. General Information. 2. Direct Competitive ELISA for Antigen Detection and Quantification. 3. Indirect Assay-Antigen Competition. 4. Indirect Competition Assay for Antibody Detection. 5. Indirect Assay Competition for Antibody-Detection Using a Single Dilution of Test Serum. 6. Overall Conclusions on Competition Assays.
Immunochemical Techniques: 1. Introduction. 2. Labeling Antibodies with Enzymes. 3. Coupling Antibodies to Alkaline Phosphatase. 4. Preparation of Immunoglobulins. 5. Immunosorbents. 6. Production of Antisera.
Index.