FluoroSpot Plus: Human IFN‑γ/IL‑2

FSP-0102-2

Publications: 49

Documents

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Ensure reproducibility when diving into a multiplex analysis.

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)

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Delivery 4-9 business days

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Complementary products

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PepPool: CEF (CD8), human

Product specifications

Intended use

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

Serum/Plasma samples

Recommendation

Product details

Product	FluoroSpot Plus: 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. Plates may be kept at room temperature.
Shelf life	At least 12 months from date of receipt.

Performance

PBMC (250,000 cells/well) were incubated overnight in the absence or presence of the CEF peptide pool (available from Mabtech, code 3616-1) and the number of cells secreting IFN-γ and/or IL-2 was analyzed by FluoroSpot. Double-secreting cells were determined as spots having the same center point in an image overlay of LED490 (IFN-γ) and LED550 (IL-2) images. Spot analysis was made with Mabtech IRIS.

Documents

Datasheet/Protocol
SDS

Product highlights

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.

Highlighted research, Published May 17, 2022

Third vaccine dose important for T cell response against Omicron

Three doses mRNA vaccine had better T cell responses than recipients of two doses. Mabtech IRIS and FluoroSpot Flex: Human IFN-γ/TNF-α/IL-2/Granzyme B were used to assess T cell immunity in mRNA vaccine recipients.

Highlighted research, Published December 16, 2021

SARS-CoV-2-specific immune responses in patients with autoimmune diseases

Patients with rheumatic diseases showed T cell responses comparable to healthy control. Patients with MS and after mRNA vaccination show different memory B cell IgG responses. SARS-CoV-2 specific immune responses in autoimmunity was analyzed using FluoroSpot, ELISpot Path, and Mabtech IRIS.

Highlighted research, Published December 2, 2021

Importance of studying SARS-CoV-2-specific memory B cells

When studying antibodies in circulation, remember that memory B cells might hide out of sight. In this study, detection of SARS-CoV-2-specific T and memory B cells 6 and 15 months after infection.

Highlighted research, Published November 24, 2021

Polyfunctional T cells with Mabtech IRIS

Study of development and persistence of virus-specific cellular and humoral immune responses, during the acute phase and two follow-ups after COVID-19 disease in hospitalized patients. Mabtech’s FluoroSpot Plus: Human IFN-γ/IL-2/TNF-α were analyzed with Mabtech IRIS.

Highlighted research, Published April 5, 2019

FluoroSpot more sensitive than flow cytometry

FluoroSpot assay detected cytokine-secreting cells even when as few as 10 transfected cells were added to the cell suspension. In contrast, at least 5 000 transfected cells were required to detect the cytokines by flow cytometry.

Highlighted research, Published May 29, 2018

FluoroSpot helps elucidate the pleiotropic effects of lenalidomide

Mabtech FluoroSpot Plus: IFN-γ/IL-2/TNF-α was utilized to show the polyfunctional profile of CD8+ T cells in patients with multiple myeloma after lenolidomide treatment.

Tutorials

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.

Publications (49)

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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