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Luke O'Neill and Andrew Bowie bring together a collection of standard and advanced methods for measuring these powerful agents. Their readily reproducible techniques range from the assay of interleukin protein and mRNA-using ELISA, FACS, and RT-PCR-to the study of interleukin signal transduction. Newer techniques are also covered, including the analysis of interleukin gene polymorphisms and the use of cDNA microarrays. Many of the assays are geared to specific pathologies, including breast cancer, depression, psoriasis, Grave's disease, migraine, and myocardial infarction. Comprehensive and highly practical, Interleukin Protocols offers biomedical investigators a stellar collection of all the major techniques needed to analyze the role of interleukins in disease, to improve diagnosis, and to foster the rapid emergence of new and more powerful therapeutics.
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M E T H O D S I N M O L E C U L A R M E D I C I N E TM Interleukin Protocols Edited by Luke A. J. O’Neill Andrew Bowie Humana Press ELISAs and Interleukin Research 3 1 ELISAs and Interleukin Research Catherine Greene 1. Introduction 1.1. Overview and Application of ELISAs ELISA (enzyme-linked immunosorbent assay) is a powerful, versatile, precise, and reliable quantitative technique for the measurement of antigens or antibodies in biologic samples. The ELISA technique is a widely used tool in biologic and biomedical research; it has been modified and adapted for multiple applications since its development almost 30 years ago (1). As the most commonly used immunoassay technique, ELISA provides the basis for numerous tests in the study of infectious diseases, epidemiology, endocrinology, and immunology. The development of specific ELISA antibodies and reagents has helped to revolutionize the field of immunology, in particular by providing a simple