FluoroSpot Plus: Human IFN‑γ/IL‑4/IL‑5

FluoroSpot Plus: Human IFN‑γ/IL‑4/IL‑5

FSP-011608-2

Publications: 1
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Ensure reproducibility when diving into a multiplex analysis.

Components

PlatePre-coated FluoroSpot plate (mAbs 1-D1K, IL4-I, and TRFK5)
Detection mAbsAnti-IFN-γ mAb (7-B6-1), BAM
Anti-IL-4 mAb (IL4-II), 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)

In stock

Delivery 4-9 business days

Shipping $0

Product specifications

Intended use

This FluoroSpot Plus kit is intended for the enumeration of cells secreting human IFN-γ, IL-4, 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-4/IL-5
ApplicationFluoroSpot
AnalyteIFN-γ, IL-4, IL-5
ReactivityHuman
SpecificityThis kit is based on matched pairs of mAbs specific for human IFN-γ, IL-4, 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.

PBMC (250,000 cells/well) were incubated overnight in the absence or presence of mixture of CEF and CEFTA peptide pools (available from Mabtech, code 3616-1, 3617-1) and anti-CD28 mAb. The number of cells secreting IFN-γ, IL-4 and 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-4) and LED640, Cy5 (IL-5). 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-4

Analyte description

Interleukin 4 (IL-4) is produced primarily by Th2 cells, mast cells, eosinophils, and basophils. IL-4 was first identified to costimulate B cell growth; the cytokine is essential for B cell secretion of IgE and augments the production of IgG1 in vivo. In addition to regulating B cell growth and immunoglobulin secretion, IL-4 also affects T cells. In vitro, IL-4 promotes T cell growth and can induce cytolytic T cell activity. IL-4 also induces the differentiation of naive CD4 T cells into Th2 cells, which are characterized by their capacity to secrete the cytokines IL-4, IL-5, and IL-10 upon activation, while simultaneously inhibiting the generation of Th1 cells.

Alternative namesInterleukin 4, IL-4, IL4, BCGF-1, BSF-1, BCGF1, BSF1
Cell typeT cell, Th2, Treg
Gene ID3565

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