Abstract
ELISpot is a highly sensitive method in immunology to enumerate cells producing a given cytokine. Cells are stimulated in a microtiter plate pre-coated with a specific anti-analyte antibody. In response to the stimulation, cells release cytokines that are bound to the anti-analyte antibody. After a washing step, which removes the cells from the wells, the location of the cytokine-releasing cell is visualized by an enzyme-labeled detection antibody and its corresponding chromogenic substrate. The end result is a set of colored spots, each of which represents an area where a cell secreting the cytokine had been located.
Here we describe the standard ELISpot protocol and a variation denominated dendritic cell (DC)-ELISpot for the detection of IFNγ-secreting cells upon stimulation with oligopeptides and protein antigens, respectively.
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Navarrete, M.A. (2015). ELISpot and DC-ELISpot Assay to Measure Frequency of Antigen-Specific IFNγ-Secreting Cells. In: Hnasko, R. (eds) ELISA. Methods in Molecular Biology, vol 1318. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2742-5_8
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DOI: https://doi.org/10.1007/978-1-4939-2742-5_8
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2741-8
Online ISBN: 978-1-4939-2742-5
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