title: ELISPOT Techniques.
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.
authors.affiliation: Department of Biology, The University of Texas at San Antonio, San Antonio, TX, USA.
authors.forename: Niannian
authors.initials: N
authors.lastname: Ji
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Forsthuber
mesh_terms: D000818:Animals; D016207:Cytokines; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods*; D006801:Humans; D051379:Mice
chemical_list: D016207:Cytokines