Detection of cytokine with enzyme-linked immunospot (ELISPOT) assay

The common method for quantitative analysis of cytokine consists of measuring cytokines produced and accumulated in the medium of short term cultures, by means of enzyme immunoassays. However, this approach provides only cumulative data, and sensitivity of the techniques used is limited, particularly in situations where relatively few cells are producing the cytokine studied. In addition, this strategy dose not permit an estimation of the frequency of the corresponding secreting cells.

The method based on immuno-enzyme technology was developed for enumerating specific antibody-secreting cells. Since its original description, the method, termed an enzyme-linked immunospot assay (ELISPOT assay), has been employed not only to detect antibody-secreting cells but also to enumerate a variety of cytokine-secreting cells. The technique is a very useful tool for quantitative analysis of cytokine at single cell level. In fact, a lot of researchers have reported significant results for cytokine research by using ELISPOT assay.

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