Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022:2386:61-79.
doi: 10.1007/978-1-0716-1771-7_5.

Detection of Cytokine-Secreting Cells by Enzyme-Linked Immunospot (ELISpot)

Bernt Axelsson  1
Affiliations

Detection of Cytokine-Secreting Cells by Enzyme-Linked Immunospot (ELISpot)

Bernt Axelsson. Methods Mol Biol. 2022.

Abstract

The enzyme-linked immunospot (ELISpot) is a highly sensitive immunoassay that measures the frequency of cytokine-secreting cells at the single-cell level. The secreted molecules are detected by using a detection antibody system similar to that used in the enzyme-linked immunosorbent assay (ELISA). The ELISpot assay is carried out in a 96-well plate and an automated ELISpot reader is used for analysis. The assay is easy to perform, robust and allows rapid analysis of a large number of samples and is not limited to measurement of cytokines; it is suitable for almost any secreted protein where single-cell analysis is of interest.

Keywords: 96-well plate; Capture antibodies; Cytokines/analytes; Detecting antibodies; Enzyme substrates; PVDF membrane.

PubMed Disclaimer

References

    1. Czerkinsky CC, Nilsson LA, Nygren H, Ouchterlony O (1983) A solid-phase enzyme-linked immunospot (ELISPOT) assay for enumeration of specific antibody-secreting cells. J Immunol Methods 65:109–121 - DOI
    1. Sedgwick JD, Holt PG (1983) A solid-phase immunoenzymatic technique for the enumeration of specific antibody-secreting cells. J Immunol Methods 57:301–309 - DOI
    1. Van de Pol MA, Lutter R, van Ree R, van der Zee JS (2012) Increase in allergen-specific IgE and ex vivo Th2 responses after a single bronchial challenge with house dust mite in allergic asthmatics. Allergy 67(1):67–73 - DOI
    1. Oseroff C, Sidney J, Tripple V, Grey H, Wood R, Broide DH, Greenbaum J, Kolla R, Peters B, Pomés A, Sette A (2012) Analysis of T-cell responses to the major allergens from German cockroach: epitope specificity and relationship to IgE production. J Immunol 189(2):679–688 - DOI
    1. Hill PC, Brookes RH, Fox A, Jackson-Sillah D, Jeffries DJ, Lugos MD, Donkor SA, Adetifa IM, de Jong BC, Aiken AM, Adegbola RA, McAdam KP (2007) Longitudinal assessment of an ELISpot test for Mycobacterium tuberculosis infection. PLoS Med 4(6):e192 - DOI

LinkOut - more resources