FluoroSpot Plus: Human IFN-γ/IL-10/IL-5

FluoroSpot Plus: Human IFN‑γ/IL‑10/IL‑5

FSP-010708-2

Publications: 8
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Components

PlatePre-coated FluoroSpot plate (mAbs 1-D1K, 9D7, and TRFK5)
Detection mAbsAnti-IFN-γ mAb (7-B6-1), BAM
Anti-IL-10 mAb (12G8), 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-10, 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-10/IL-5
ApplicationFluoroSpot
AnalyteIFN-γ, IL-10, IL-5
ReactivityHuman
Specificity

This kit is based on matched pairs of mAbs specific for human IFN-γ, IL-10, 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.
PBMC (250,000 cells/well) were incubated in the absence or presence of tetanus toxin and anti-CD28 monoclonal antibody for 48 hours and the number of cells secreting IFN-γ, IL-10 and/or IL-5 was analyzed by FluoroSpot. Double- and triple secreting cells were determined as spots having the same position (centre point) in an image overlay of IFN-γ, IL-10 and IL-5 images. 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-10

Analyte description

Interleukin 10 (IL-10) is a pleiotropic cytokine produced by activated T cells and monocytes. It can modulate the functions of immune responses in both a stimulatory and an inhibitory way. IL-10 is thereby involved in the regulation of immune reactions and inflammatory responses.

Alternative namesInterleukin-10, IL-10, IL10, CSIF, GVHDS, IL-10A, TGIF
Cell typeT cell, Treg, Monocyte/MΦ, mDC
Gene ID3586

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