ELISpot and DC-ELISpot Assay to Measure Frequency of Antigen-Specific IFN gamma-Secreting Cells

Navarrete, Marcelo A.; Hnasko, R


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 enzymelabeled 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 IFN gamma-secreting cells upon stimulation with oligopeptides and protein antigens, respectively.

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Título según WOS: ID WOS:000375613000009 Not found in local WOS DB
Volumen: 1318
Editorial: Humana Press, Inc.
Fecha de publicación: 2015
Página de inicio: 79
Página final: 86


Notas: ISI