FluoroSpot Plus: Human IFN‑γ/IL‑5/TNF‑α

FluoroSpot Plus: Human IFN‑γ/IL‑5/TNF‑α

FSP-010809-2

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

Components

PlatePre-coated FluoroSpot plate (mAbs 1-D1K, TRFK5, and MT25C5)
Detection mAbsAnti-IFN-γ mAb (7-B6-1), BAM
Anti-IL-5 mAb (5A10), biotin
Anti-TNF-α mAb (MT20D9), 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-5, and TNF-α 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-5/TNF-α
ApplicationFluoroSpot
AnalyteIFN-γ, IL-5, TNF-α
ReactivityHuman
SpecificityThis kit is based on matched pairs of mAbs specific for human IFN-γ, IL-5, and TNF-α.

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.

Human PBMC (250,000 cells/well) were stimulated with a mixture of CEF extended, CMV, and CEFTA peptide pools (Mabtech) and anti-CD28 for 42 h and the number of cells secreting IFN-γ, IL-5 and TNF-α were analyzed by FluoroSpot using 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-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

TNF-α

Analyte description

Tumor necrosis factor (TNF), also known as TNF-α, is produced by many different cell types, e.g., monocytes, macrophages, T cells, and B cells. Among the many effects of TNF-α are protection against bacterial infection, cell growth modulation, immune system regulation, and involvement in septic shock.

Alternative namesTumor necrosis factor-α, TNF-α, TNF-alpha, TNF-a, TNFa, Tumor necrosis factor-alpha, TNF, DIF-alpha, TNFA, TNFSF2, TNLG1F
Cell typeT cell, Tc, Th1, Th2, Th17, Tfh, Monocyte/MΦ
Gene ID7124