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

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 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.

Tutorial, Published November 24, 2022

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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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