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ELISPOT Techniques

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Multiple Sclerosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1304))

Abstract

The enzyme-linked immunospot (ELISPOT) assay is a widely used method for enumerating antigen-specific cytokine-producing or antibody-secreting immune cells. It is one of the most effective immunological and diagnostic approaches to detect and quantify low-frequency cytokine- or antibody-producing cells in human and animal tissues, such as peripheral blood, lymph nodes, and spleen. Detection and quantification of specific cytokine-producing cells by the ELISPOT assay is based on the formation of visible spots at the site of cytokine release by the cells under investigation (e.g., T cells) using pairs of different capture and detection antibodies under optimized conditions.

Here we focus mainly on practical, optimized protocols for cytokine ELISPOT assays for detection of mouse and human cytokine-producing immune cells (e.g., peripheral blood mononuclear cells, PBMC), including suggestions for trouble-shooting and optimizing steps for problematic tissue samples.

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Acknowledgement

The authors would like to thank Dr. Neal Guentzel at the University of Texas at San Antonio for his valuable comments and suggestions during revision.

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Correspondence to Thomas G. Forsthuber .

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© 2014 Springer Science+Business Media New York

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Ji, N., Forsthuber, T.G. (2014). ELISPOT Techniques. In: Weissert, R. (eds) Multiple Sclerosis. Methods in Molecular Biology, vol 1304. Humana Press, New York, NY. https://doi.org/10.1007/7651_2014_111

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  • DOI: https://doi.org/10.1007/7651_2014_111

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2629-9

  • Online ISBN: 978-1-4939-2630-5

  • eBook Packages: Springer Protocols

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