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AID EliSpot Assay method

Detection of secreted cytokines

Picturing single T cells and its cytokine production

The assay is performed in a multitestplate, in which the membrane in each well is coated with specific antibodies against the cytokines to detect.

To activate the T cells, antigen or mitogen solution is mixed in each well with the isolated PBMCs.

The antigen-specific, activated T cells produce cytokines which are bound by the coated antibodies.

The secrected cytokines are detected by a secondary, conjugated antibody and a following chromogen.

The resulting spots in the wells are corresponding to the number of antigen-specific T cells.