FluoroSpot Plus: Human IFN‑γ/IL‑17A/IL‑5

Human PBMC incubated in the absence or presence of TT and anti-CD28 mAb and the number of cells secreting IFN-γ, IL-17A and/or IL-5 was analyzed by FluoroSpot. Spot analysis was made with Mabtech IRIS.

FluoroSpot Plus: Human IFN‑γ/IL‑17A/IL‑5

FSP-010308-2

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Components

PlatePre-coated FluoroSpot plate (mAbs 1-D1K, MT44.6, and TRFK5)
Detection mAbsAnti-IFN-γ mAb (7-B6-1), BAM
Anti-IL-17A mAb (MT504), biotin
Anti-IL-5 mAb (5A10), 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-17A, and IL-5 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-17A/IL-5
ApplicationFluoroSpot
AnalyteIFN-γ, IL-17A, IL-5
ReactivityHuman
SpecificityThis kit is based on matched pairs of mAbs specific for human IFN-γ, IL-17A, and IL-5.

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 incubated in the absence or presence of TT and anti-CD28 mAb and the number of cells secreting IFN-γ, IL-17A and/or IL-5 was analyzed by FluoroSpot. Spot analysis was made with Mabtech IRIS.
Human PBMC were incubated in the absence or presence of TT and anti- CD28 monoclonal antibody for 48 hours and the number of cells secreting IFN-γ, IL-17A and/or IL-5 was analyzed by FluoroSpot. Single cytokine secreting cells as well as double- and triple-secreting cells were determined. Spot center was used to identify spots from double- and triple-secreting cells, visualized in an image overlay of LED490, FITC (IFN-γ) and LED550, Cy3 (IL-22) and LED640, Cy5 (IL-17A). Spot analysis was made with Mabtech IRIS.
Human PBMC incubated in the absence or presence of TT and anti-CD28 mAb and the number of cells secreting IFN-γ, IL-17A and/or IL-5 was analyzed by FluoroSpot. 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-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

IL-5

Analyte description

Interleukin 5 (IL-5) is a pleiotropic cytokine primarily produced by T and NK cells.  IL-5 specifically controls the survival, differentiation, and chemotaxis of eosinophils. But initially, IL-5 was characterized by its ability to support the growth and terminal differentiation of B cells.

Alternative namesInterleukin-5, IL-5, IL5, BCGF-2, EDF, TRF
Cell typeT cell, Th2, NK cell
Gene ID3567
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