FluoroSpot Plus: Human IFN-γ/TNF-α

FluoroSpot Plus: Human IFN‑γ/TNF‑α

FSP-0109-2

Publications: 3
Documents
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Ensure reproducibility when diving into a multiplex analysis.

Components

PlatePre-coated FluoroSpot plate (mAbs 1-D1K and MT25C5)
Detection mAbsAnti-IFN-γ mAb (7-B6-1), BAM
Anti-TNF-α mAb (MT20D9), biotin
Fluorophore conjugatesAnti-BAM mAb, 490
SA-550
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 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/TNF-α
ApplicationFluoroSpot
AnalyteIFN-γ, TNF-α
ReactivityHuman
Specificity

This kit is based on matched pairs of mAbs specific for human IFN-γ 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 incubated overnight in the absence or presence of the CEF peptide pool and anti-CD28 antibody (both available from Mabtech) and the number of cells secreting IFN-γ and/or TNF-α were analyzed by FluoroSpot. Double-secreting cells were determined as spots having the same center point in an image overlay of LED490, FITC (IFN-γ) and LED550, Cy3 (TNF-α) images. Spot analysis was made with Mabtech IRIS.

Highlighted research, Published October 11, 2024

Immunoassays for vaccine research
Explore the critical role of immunoassays like ELISpot and FluoroSpot in vaccine development, from evaluating immune responses to tracking vaccine efficacy and potency. These assays provide sensitive, single-cell insights for advancing research and development of viral, bacterial, and cancer vaccines.
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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

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
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