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The enzyme-linked immunospot (ELISPOT) assay is designed to enumerate cytokine producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in vitro manipulations allowing cytokine production analysis as close as possible to in vivo conditions. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or pathological state. To perform the ELISPOT assay, cytokine-specific antibodies are first attached to the surface of a plastic well. Next, cytokine-secreting cells (i.e. activated T cells) are added to the wells. Cytokine that is secreted by these cells are bound by cytokine-specific antibodies. Finally, cytokine-secreting cells are revealed by a second cytokine-specific antibody that is enzyme-labeled; after a reaction with the appropriate substrate, each cell secreting cytokine is revealed as a spot.
For more detailed information about the ELISPOT assay and how it is performed, please refer to this .pdf or visit Cell Sciences.
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