ELISPOT Techniques

  • Niannian Ji
  • Thomas G. ForsthuberEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1304)


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.


ELISPOT Antigen recall Cytokine Antibody Immune cells T cells PBMCs 



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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of BiologyThe University of Texas at San AntonioSan AntonioUSA

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