Introduction to the ELISpot experimental operation process

Introduction to the ELISpot experimental operation process

The full name of ELISPOT is enzyme -linked immunospot assay, English: Enzyme-linked Immunospot Assay . It combines cell culture technology with enzyme-linked immunosorbent assay ( ELISA ) to detect cytokine secretion at the single cell level. The technical principle is summarized in one sentence: the cytokine secreted by the cells in the culture is captured by the antibody, and is expressed by enzyme-linked spot color development.

Cytokine-specific monoclonal antibodies were coated onto ethanol treated PVDF membrane plates . For specific board processing and coating methods, please refer to :
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The cells are added to the wells of the enzyme, and different groups are given different stimulants or no stimulants .   For specific cell processing methods, please refer to :
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The cytokine secreted by the cell binds to the coated antibody .

After the excess cells are removed and washed , the biotin-labeled detection antibody is added to the well-labeled wells .

Addition of a reciprocin-labeled enzyme conjugate

Add substrate
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Spots were read and analyzed using an enzyme-linked immunospot reader . Source of information :
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