R&D Systems Dual-Color FluoroSpot Kits utilize the ELISpot assay principle to simultaneously detect individual cells that secrete either one or both analytes specified in the kit. These assays are highly sensitive and can quantitate actively secreting cells even when cell frequencies fall below 1 in 100,000. Unlike other methods that assess antigen-specific immune cell responses, this assay does not require prior in vitro expansion of cells and is suitable for high-throughput analysis. The Dual-Color FluoroSpot Kits utilize two spectrally distinct NorthernLights™ fluorescent probes to detect the analyte-secreting cells. All complete FluoroSpot Kits are complete ready-to-run assays and require no further development or refinement.
Detect and quantitate individual cells secreting specific analytes
High sensitivity – FluoroSpot kits can measure responses with frequencies below 1 in 100,000 cells
No in vitro expansion of cells required
Amenable to high-throughput
R&D Systems Dual-Color FluoroSpot Kits employ antibody-coated, membrane-backed microplates to identify secreting cells. The immobilized capture antibodies bind to cytokines that have been secreted in their vicinity, and two spectrally distinct NorthernLights™ fluorescent probes are used to visualize the secreted proteins. Fluorescent spots appear at the sites of cytokine localization, with each individual spot representing an individual cytokine-secreting cell.
Dual-Color Kit Contents
PVDF-backed 96-well Microplate
NorthernLights Fluorescence Enhancer
Detection of IFN-gamma and IL-2 Secretion by Human PBMCs using the Dual-Color FluoroSpot Kit
Human peripheral blood mononuclear cells (PBMCs) were stimulated overnight with PMA/Ca2+ ionomycin and assessed for IFN-gamma and IL-2 secretion using the Human IFN-gamma/IL-2 Dual-Color FluoroSpot Kit (Catalog # ELD4506NL). Spots of IFN-gamma secretion were visualized in green, IL-2 secretion in red, and secretion of both cytokines in yellow.