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 IFN-γ/IL-10/IL-2 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 and includes low fluorescent plates pre-coated with monoclonal capture antibodies, monoclonal detection antibodies, secondary reagents conjugated to fluorophores, a fluorescence enhancer and an anti-CD28 monoclonal antibody for co-stimulation. A polyclonal activator for human T cells (mAb CD3-2) is supplied as a positive control for cytokine secretion.

Pre-coated plates enable a more rapid assay with minimized variability.

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.

IFN-γInterferon-γ (IFN-γ, also known as IFN-gamma, IFN-g, IFNg and Interferon-gamma) is produced mainly by activated T cells and NK cells. It is a proinflammatory cytokine that activates macrophages and endothelial cells, but it also regulates immune responses by effecting APC and T and B cells. Production of IFN-γ by helper T cells as well as cytotoxic T cells is a hallmark of the TH1-type phenotype, thus, high-level production of IFN-γ is typically associated with effective host defense against intracellular pathogens.

IL-10Interleukin-10 (IL-10, IL10) is a pleiotropic cytokine that is produced by activated T cells and monocytes. It can modulate the functions of immune responses in both a stimulatory and an inhibitory way. IL-10 is thereby involved in the regulation of immune reactions and inflammatory responses.

IL-2Interleukin-2 (IL-2, IL2) is produced exclusively by activated T cells. IL-2 promotes the proliferation, differentiation, and survival of antigen-activated T cells and NK-cells.

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