FluoroSpot Plus: Human IL-22/IFN-γ/IL-5/IL-17A

FluoroSpot Plus: Human IL‑22/IFN‑γ/IL‑5/IL‑17A

FSP-18010803-2

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Components

PlatePre-coated FluoroSpot plate (mAbs MT12A3, 1-D1K, TRFK5, and MT44.6)
Detection mAbsAnti-IL-22 mAb (MT7B27), DIG
Anti-IFN-γ mAb (7-B6-1), BAM
Anti-IL-5 mAb (5A10), biotin
Anti-IL-17A mAb (MT504), WASP
Fluorophore conjugatesAnti-DIG mAb, 380
Anti-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 IL-22, IFN-γ, IL-5, and IL-17A using the FluoroSpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

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Product details

ProductFluoroSpot Plus: Human IL-22/IFN-γ/IL-5/IL-17A
ApplicationFluoroSpot
AnalyteIL-22, IFN-γ, IL-5, IL-17A
ReactivityHuman
Specificity

This kit is based on matched pairs of mAbs specific for human IL-22, IFN-γ, IL-5, 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 PPD and anti-CD28 antibody and the number of cells secreting IL-22, IFN-γ, IL-5 and IL-17A was analyzed by FluoroSpot. Single cytokine secreting cells as well as cells secreting two, three or four analytes were determined. Spot center was used to identify spots from cells secreting multiple analytes, visualized in an image overlay of LED380 (IL-22), LED490 (IFN-γ) and LED550, (IL-5) and LED640, Cy5 (IL-17A). Spot analysis was made with Mabtech IRIS.

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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

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-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

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