Handbook of ELISPOT Methods and Protocols

Although the ELISPOT (enzyme-linked immunospot) assay is widely used in many fields of biomedical research, producing recognizable and quantifiable spots may be very difficult not only for beginners, but also for experienced researchers. In Handbook of ELISPOT: Methods and Protocols, the first book dedicated entirely to the ELISPOT, recognized experts with first-hand experience detail how to design, perform, and analyze these assays. The readily reproducible techniques they provide cover a wide variety of topics, including the use of membrane-backed plates, the standardization and validation procedures, the removal of cells from ELISPOT plates, cell separation techniques, and the quantification of ELISPOT data. There are also numerous ELISPOT applications involving animal models, human cells, measles, multiple sclerosis, immune responses, multicytokine detection systems, and immunocytochemistry. Highlights include dual-color and multiplex ELISPOT assays, use of the ELISPOT assay on feline lymphocytes, standardization of the ELISPOT procedure, and combining the ELISPOT assay with immunohistochemistry. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and highly practical, Handbook of ELISPOT: Methods and Protocols offers both novice and experienced researchers a wealth of easy-to-understand information that will clarify the technically challenging aspects of ELISPOT and open the door to its wider successful use.