Main description:

This volume is a practical biochemical guide to the Enzyme-Linked Immunosorbent Assay (ELISA), used to detect a target substance in a liquid sample. The ELISA is an important and widely used diagnostic tool in medicine, animal health, botany and quality assurance processes in food and beverage production. An introductory chapter orients the reader on the basic structure and function of immunoglobulins and their fragments while subsequent chapters outline the methodology to generate monoclonal antibodies using hybridoma technology and the general methods used to purify antibodies. Multiple chapters demonstrate how to creatively use the properties of the antibody to identify, localize and quantify target analytes to answer questions and resolve problems. The reader will learn how to use a variety of immunoassay strategies, reporters and detection systems that will undoubtedly facilitate their efforts to gain answers to their own questions. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, ELISA: Methods and Protocols seeks to provide both professionals and novices with the technical information necessary for the reader to successfully use the immunoassay as part of the discovery process.

Contents:

1. The Biochemical Properties of Antibodies and their Fragments

Robert M. Hnasko

2. Hybridoma Technology

Robert M. Hnasko and Larry H. Stanker

3. Affinity Purification of Antibodies

Robert M. Hnasko and Jeffery A. McGarvey

4. Bio conjugation of Antibodies to Horseradish Peroxidase (HRP)

Robert M. Hnasko

5. Indirect ELISA

Alice V. Lin

6. Direct ELISA

Alice V. Lin

7. A Double-Sandwich ELISA for Identification of Monoclonal Antibodies Suitable for Sandwich Immunoassays

Larry H. Stanker and Robert M. Hnasko

8. ELISpot and DC-ELISspot Assay to Measure Frequency of Antigen-Specific IFNg Secreting Cells

Marcelo A. Navarrete

9. The Western Blot

Thomas S. Hnasko and Robert M. Hnasko

10. Development of Flow Through Assay for Detection of Antibodies to Taenia solium Using Protein-A Colloidal Gold Conjugate as a Probe

Chennuru Sreedevi and Panduranga Rao Pavuluri

11. Multiplexed Microsphere Suspension Array-Based Immunoassays

Andrew Lin, Alexandra Salvador, and J. Mark Carter

12. Multiplex Immunoassay: A Planar Array on a Chip using the MagArray (TM) Technology

Laurie M. Clotilde, Heng Yu, and M. Luis Carbonell

13. Lateral Flow Immunoassay

Kathryn H. Ching

14. Immuno-PCR Assay for Sensitive Detection of Proteins in Real Time

Xiaohua He and Stephanie A. Patfield

15. In-situ Proximity Ligation Assay (PLA)

Sonchita Bagchi, Robert Fredriksson, and Asa Wallen-Mackenzie

16. Tyramide Signal Amplification for Immunofluorescent Enhancement

Lauren Faget and Thomas S. Hnasko

17. Correlative microscopy for Localization of Proteins In Situ: Pre-Embedding Immuno-Electron Microscopy using FluoroNanogold, Gold Enhancement and Low-Temperature Resin

Daniela Boassa

18. Multiplex ELISA using Oligonucleotide Tethered Antibodies

Robert S. Matson

19. Gas Plasma Surface Chemistry for Biological Assays

Khoren Sahagian and Mikki Larner