FluoroSpot Plus: Human IFN-γ/IL-4/TNF-α

FluoroSpot Plus: Human IFN‑γ/IL‑4/TNF‑α

FSP-011609-2

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Components

PlatePre-coated FluoroSpot plate (mAbs 1-D1K, IL4-I, and MT25C5)
Detection mAbsAnti-IFN-γ mAb (7-B6-1), BAM
Anti-IL-4 mAb (IL4-II), 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-4, 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-4/TNF-α
ApplicationFluoroSpot
AnalyteIFN-γ, IL-4, TNF-α
ReactivityHuman
Specificity

This kit is based on matched pairs of mAbs specific for human IFN-γ, IL-4, and TNF-α.

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.
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-4 and TNF-α were analyzed by FluoroSpot using 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-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

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