FluoroSpot Plus: Human IFN-γ/IL-22/IL-17A

FluoroSpot Plus: Human IFN-γ/IL-22/IL-17A

FSP-011803-2

Publications: 6
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Components

Detection mAbsAnti-IFN-γ mAb (7-B6-1), BAM
Anti-IL-22 mAb (MT7B27), biotin
Anti-IL-17A mAb (MT504), WASP
Fluorophore conjugatesAnti-BAM mAb, 490
SA-550
Anti-WASP mAb, 640
Positive controlAnti-CD3 mAb (CD3-2)
Co-stimulusAnti-CD28 mAb (CD28-A)
Buffer/SolutionFluoroSpot enhancer
PlatePre-coated FluoroSpot plate (mAbs 1-D1K, MT12A3, and MT44.6)

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Product specifications

Intended use

This FluoroSpot Plus kit is intended for the enumeration of cells secreting human IFN-γ, IL-22, 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-22/IL-17A
ApplicationFluoroSpot
AnalyteIFN-γ, IL-22, IL-17A
ReactivityHuman
Specificity

This kit is based on matched pairs of mAbs specific for human IFN-γ, IL-22, and IL-17A.

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.
PBMC (250,000 cells/well) were incubated for 48 hours in the absence or presence of the CEF peptide pool and anti-CD28 antibody (both available from Mabtech) and the number of cells secreting IFN-γ, IL-22 and IL-17A 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.
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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-22

Analyte description

The cytokine interleukin 22 (IL-22) is mainly produced by activated CD4 T helper 17 (Th17) and T helper 22 (Th22) cells. The IL-22 receptor is expressed on non-immune cells, particularly epithelial cells and keratinocytes, and IL-22 promotes innate immune responses versus bacterial infections in these target cells. In addition, Th22 cells have been shown to play a role in the pathophysiology of several human skin diseases.

Alternative namesInterleukin-22, IL-22, IL22, IL-21, IL-D110, IL-TIF, ILTIF, TIFIL-23, TIFa, zcyto18
Cell typeT cell, Th1, Th17, Tfh
Gene ID50616

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
Cell typeTh17