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

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

FSP-414443-2

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Components

PlatePre-coated FluoroSpot plate (mAbs AN18, IL17-I, and TRFK5)
Detection mAbsAnti-IFN-γ mAb (R4-6A2), BAM
Anti-IL-17A mAb (MT2270), biotin
Anti-IL-5 mAb (TRFK4), WASP
Fluorophore conjugatesAnti-BAM mAb, 490
SA-550
Anti-WASP mAb, 640
Buffer/SolutionFluoroSpot enhancer
Co-stimulusAnti-CD28 mAb

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Delivery within 3 weeks

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Product specifications

Intended use

This FluoroSpot Plus kit is intended for the enumeration of cells secreting mouse 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: Mouse IFN-γ/IL-17A/IL-5
ApplicationFluoroSpot
AnalyteIFN-γ, IL-17A, IL-5
ReactivityMouse
Specificity

This kit is based on matched pairs of mAbs specific for mouse IFN-γ, IL-17A, and IL-5.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store 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.
Mouse spleen cells were incubated overnight in the absence or presence of Concanavalin A (ConA) and anti-CD28 antibody. The number of cells secreting IFN-γ, IL-17A and/or IL-5 was analyzed by FluoroSpot. Double- and triple secreting cells were determined as spots having the same center point in an image overlay. Spot analysis was made with Mabtech IRIS.

Tutorial, Published November 24, 2022

Cell stimulation in ELISpot and FluoroSpot
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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, If2f, Ifg
Cell typeT cell, Tc, Th1, NK cell
Gene ID15978

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
Cell typeTh17
Gene ID16171

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, Il-5
Cell typeT cell, Th2, NK cell
Gene ID16191