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 human IL-1β/IL-6/TNF-α FluoroSpot kit is perfect for users who wish to have a convenient assay with minimized assay variability. It enables enumeration of cells secreting one, two or three analytes. The kit includes low fluorescent plates pre-coated with monoclonal capture antibodies, monoclonal detection antibodies, secondary reagents conjugated to fluorophores and a fluorescence enhancer. LPS is included as positive control stimulus.

The assay is recommended for the analysis of macrophages, monocytes and dendritic cells. These kits have not been evaluated for T-cells.

For analysis, we recommend the use of an automated FluoroSpot reader equipped with filters for excitation 490 nm/emission 510 nm (LED490, FITC) for 490-reagent detection, excitation 550 nm/emission 570 nm (LED550, Cy3) for 550-reagent detection and excitation 640 nm/ emission 660 nm (LED640, Cy5) for 640-reagent detection. Filters should have high specificity to avoid bleed-through artifacts.

IL-1βInterleukin-1ß (IL-1ß or IL-1F2, also known as Interleukin-1beta, IL-1 beta, IL1b and Interleukin-1 beta) is a proinflammatory cytokine and inducer of acute phase responses. IL-1ß is produced primarily by monocytes, macrophages, and dendritic cells after induction by microbes.

IL-6Interleukin 6 (IL-6, IL6) is a pleiotropic cytokine that is produced by many different cell types and plays a role in a wide range of functions, such as immune responses, acute-phase reactions, and hematopoiesis. Among other things it augments antibody production from activated B cells in vitro.

TNF-αTumor necrosis factor-α (TNF-α, also known as TNF-alpha, TNF-a, TNFa, and Tumor necrosis factor-alpha) is produced by many different cell types e.g. monocytes, macrophages, T cells and B cells. Among the many effects of TNF-α, are protection against bacterial infection, cell growth modulation, immune system regulation and involvement in septic shock.

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