FluoroSpot Path: CMV Human IFN-γ/IL-2

FluoroSpot Path: CMV Human IFN-γ/IL-2

FSP-0102-P2-1

Publications: 46
Documents
Questions?
Paves your way to functional analysis of a specific research question.

Components

Peptide poolCMV (CD4 and CD8), human
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

In stock

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 CMV peptides using the FluoroSpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

Peptide pool composition
The CMV peptide pool contains 42 peptides derived from CMV. The antigens represented in the peptide pool are pp50, pp65, IE1, IE2, and Envelope glycoprotein B. Of the included peptides, 28 are MHC class I restricted and 14 are MHC class II restricted.

Recommendation

Product details

ProductFluoroSpot Path: CMV 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. The peptide pool 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) were incubated overnight in the absence or presence of the CMV peptide pool for human CD4 and CD8 T cells (Mabtech) and 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

Tutorial, Published November 24, 2022

Cell stimulation in ELISpot and FluoroSpot
In this feature, we’ll walk you through the optimal use of different polyclonal and antigen-specific stimuli.

Tutorial, Published September 2, 2022

Isolation, freezing, and thawing of PBMCs
This is a protocol on how to best isolate, freeze, and thaw PBMCs.
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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