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The FluoroSpot assay is an inventive development of the ELISpot assay and uses fluorescent detection instead of colorimetric. This allows for simultaneous detection of several analytes, which is a major advantage. The monkey IFN-γ/IL-2 FluoroSpot kit enables convenient enumeration of cells secreting either one or both analytes. The kit contains low fluorescent plates, matched quantities of monoclonal capture and detection antibodies, secondary reagents conjugated to fluorophores, a fluorescence enhancer and an anti-CD28 monoclonal antibody for co-stimulation. The included anti-CD3 mAb can be used to induce cytokine secretion and is dependent on number, nature and viability of accessory cells. The antibody cross-reacts with CD3 on cynomolgus and rhesus macaque T cells.