FluoroSpot Path: TB (ESAT-6+CFP-10+EspC) Human IFN-γ/IL-2

FluoroSpot Path: TB (ESAT-6+CFP-10+EspC) Human IFN-γ/IL-2

FSP-0102-P4-1

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Components

Peptide poolsTB (ESAT-6), scanning
TB (CFP-10), scanning
TB (EspC), scanning
Detection mAbsAnti-IFN-γ mAb (7-B6-1), BAM
Anti-IL-2 mAb (MT8G10), biotin
Fluorophore conjugatesAnti-BAM mAb, 490
SA-550
Positive controlAnti-CD3 mAb (CD3-2)
Co-stimulusAnti-CD28 mAb (CD28-A)
PlatePre-coated FluoroSpot plate (mAbs 1-D1K and MT2A91/2C95)
Buffer/SolutionFluoroSpot enhancer

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Delivery 4-9 business days

Shipping $0

Product specifications

Intended use

This FluoroSpot Path kit is intended for the enumeration of cells secreting human IFN-γ and IL-2 in response to TB peptides using the FluoroSpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

Peptide pool composition
The scanning pools consist of synthetic peptides of 15-mers overlapping with 11 amino acids, covering the complete proteins. The ESAT-6 scanning pool is derived from the early secretory antigenic target protein and contains 21 peptides. The CFP-10 scanning pool is derived from the culture filtrate protein and contains 23 peptides. Finally, the EspC scanning pool is derived from the ESX-1 secretion-associated protein and contains 23 peptides.

Recommendation

Product details

ProductFluoroSpot Path: TB (ESAT-6+CFP-10+EspC) Human IFN-γ/IL-2
ApplicationFluoroSpot
AnalyteIFN-γ, IL-2
ReactivityHuman
Specificity

This kit is based on matched pairs of mAbs specific for human IFN-γ and IL-2.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store antibodies, fluorophore conjugates, and FluoroSpot enhancer at 4-8 °C upon receipt. Peptide pools should be stored at -20 °C or below. Plates may be kept at room temperature.

Shelf lifeAt least 12 months from date of receipt.
Human PBMC (250,000 cells/well) from an individual with latent tuberculosis were incubated overnight in the absence or presence of the ESAT-6 and CFP-10 scanning pools (Mabtech). The number of cells secreting IFN-γ and/or IL-2 was analyzed by FluoroSpot. Single cytokine secreting cells as well as double-secreting cells were determined. Spot center was used to identify spots from double-secreting cells, visualized in an image overlay of LED490, FITC (IFN-γ) and LED550,Cy3 (IL-2). Spot analysis was made with Mabtech IRIS.
Datasheet/Protocol
SDS

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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-2

Analyte description

Interleukin 2 (IL-2) is predominantly produced by activated T cells. IL-2 promotes the proliferation, differentiation, and survival of antigen-activated T cells and NK cells. After antigen stimulation, the cytokine also promotes the differentiation of T cells into effector and memory T cells. Therefore IL-2 plays an important role in the body's response to infection. IL-2 is also key to tolerance, as it promotes the differentiation of certain immature T cells into regulatory T cells in the thymus, ultimately preventing autoimmune diseases. 

Alternative namesInterleukin-2, Interleukin 2, IL-2, IL2, T cell growth factor, TCGF, lymphokine
Cell typeT cell, Th1, Th2, Th17, Tfh
Gene ID3558