FluoroSpot Plus: Human IFN‑γ/IL‑10/IL‑17A

FluoroSpot Plus: Human IFN‑γ/IL‑10/IL‑17A

FSP-010703-2

Publications: 5
Documents
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Ensure reproducibility when diving into a multiplex analysis.

Components

PlatePre-coated FluoroSpot plate (mAbs 1-D1K, 9D7, and MT44.6)
Detection mAbsAnti-IFN-γ mAb (7-B6-1), BAM
Anti-IL-10 mAb (12G8), biotin
Anti-IL-17A mAb (MT504), WASP
Fluorophore conjugatesAnti-BAM mAb, 490
SA-550
Anti-WASP mAb, 640
Buffer/SolutionFluoroSpot enhancer
Positive controlAnti-CD3 mAb (CD3-2)
Co-stimulusAnti-CD28 mAb (CD28-A)

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Product specifications

Intended use

This FluoroSpot Plus kit is intended for the enumeration of cells secreting human IFN-γ, IL-10, and IL-17A using the FluoroSpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

Recommendation

Product details

ProductFluoroSpot Plus: Human IFN-γ/IL-10/IL-17A
ApplicationFluoroSpot
AnalyteIFN-γ, IL-10, IL-17A
ReactivityHuman
SpecificityThis kit is based on matched pairs of mAbs specific for human IFN-γ, IL-10, and IL-17A.

Shipping and Storage

ShippingShipped at ambient temperature.
StorageStore antibodies, fluorophore conjugates, and FluoroSpot enhancer at 4-8 °C upon receipt. Plates may be kept at room temperature.
Shelf lifeAt least 12 months from date of receipt.
Human PBMC (250,000 cells/well) were incubated with CEF peptide pool and anti-CD28 antibody for 48 hours and the number of cells secreting IFN-γ, IL-10 and IL-17A were analyzed by FluoroSpot. Single cytokine secreting cells as well as double- and triple-secreting cells were determined. Spot position was used to identify spots from double- and triple-secreting cells, visualized in an image overlay of FITC (IFN-γ) and Cy3 (IL-10) and Cy5 (IL-17A). Spot analysis was made with Mabtech IRIS.

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IFN-γ

Analyte description

Interferon-γ (IFN-γ) is the only type II interferon. This proinflammatory cytokine is secreted by activated T cells and NK cells. It activates macrophages and endothelial cells and regulates immune responses by affecting APCs, T cells, and B cells. Production of IFN-γ by helper T cells and 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.

Alternative namesInterferon-γ, Interferon-gamma, IFN-γ, IFN-gamma, IFN-g, IFNg, IFG, IFI, IMD69
Cell typeT cell, Tc, Th1, NK cell
Gene ID3458

IL-10

Analyte description

Interleukin 10 (IL-10) is a pleiotropic cytokine 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.

Alternative namesInterleukin-10, IL-10, IL10, CSIF, GVHDS, IL-10A, TGIF
Cell typeT cell, Treg, Monocyte/MΦ, mDC
Gene ID3586

IL-17A

Analyte description

Interleukin 17A (IL-17A) is a potent proinflammatory cytokine produced by activated Th17 (T helper 17) cells and certain cells belonging to the innate immune system. In mice, IL-17 has also been shown to be produced by activated CD8 T cells and γδ T cells. Th17 cells play an important role in autoimmune diseases and protection against bacteria and fungi. IL-17A acts on a broad range of cell types to induce the expression of cytokines, chemokines, and metalloproteinases. As a result, secretion of IL-17A promotes inflammatory responses, which leads to the recruitment of neutrophils, enhancement of antibody production, and activation of T cells. Increased expression of IL-17A is seen in autoimmune diseases such as multiple sclerosis and rheumatoid arthritis. It is also associated with asthma, psoriasis, cancer, and transplant rejection.

Alternative namesInterleukin 17A, IL-17A, IL17A, CTLA-8, CTLA8, IL-17, IL17, ILA17
Cell typeTh17
Gene ID3605
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