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 dark Spots in the Wells are corresponding to the Nummer of antigen-specific T-cells.