FluoroSpot Path: CMV Human IFN-γ/IL-2

FluoroSpot Path: CMV Human IFN‑γ/IL‑2

FSP-0102-P2-1

Publications: 1

Documents

Questions?

Paves your way to functional analysis of a specific research question.

Components

Plate	Pre-coated FluoroSpot plate (mAbs 1-D1K and MT2A91/2C95)
Detection mAbs	Anti-IFN-γ mAb (7-B6-1), BAM
	Anti-IL-2 mAb (MT8G10), biotin
Fluorophore conjugates	Anti-BAM mAb, 490
	SA-550
Buffer/Solution	FluoroSpot enhancer
Positive control	Anti-CD3 mAb (CD3-2)
Co-stimulus	Anti-CD28 mAb (CD28-A)
Peptide pool	CMV (CD4 and CD8), human

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

Product	FluoroSpot Path: CMV Human IFN-γ/IL-2
Application	FluoroSpot
Analyte	IFN-γ, IL-2
Reactivity	Human
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 life	At least 12 months from date of receipt.

Performance

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.

Documents

Datasheet/Protocol
SDS

Tutorials

Tutorial, Published November 24, 2022

Cell stimulation in ELISpot and FluoroSpot

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Tutorial, Published September 2, 2022

Isolation, freezing, and thawing of PBMCs

This is a protocol on how to best isolate, freeze, and thaw PBMCs.

Publications (1)

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Analyte information

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 names	Interferon-γ, Interferon-gamma, IFN-γ, IFN-gamma, IFN-g, IFNg, IFG, IFI, IMD69
Cell type	T cell, Tc, Th1, NK cell
Gene ID	3458

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 names	Interleukin-2, Interleukin 2, IL-2, IL2, T cell growth factor, TCGF, lymphokine
Cell type	T cell, Th1, Th2, Th17, Tfh
Gene ID	3558