FluoroSpot Path: SARS‑CoV‑2 (S1scan+SNMO) Human IFN‑γ/IL‑2

FSP-0102-P1-1

Publications: 17

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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 pools	SARS-CoV-2 (S1), scanning
	SARS-CoV-2 (SNMO), human

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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 SARS-CoV-2 (S1 scan+SNMO) peptides using the FluoroSpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

Peptide pool composition
The SARS-CoV-2 S1 scanning pool contains 166 peptides from the human SARS-CoV-2 virus. The peptides are 15-mers overlapping with 11 amino acids, covering the S1 domain of the spike protein (amino acid 13-685). The pool is supplied in two vials: pool 1 SARS-CoV-2 S1 peptides 1-83 (83 peptides) and pool 2 SARS-CoV-2 S1 peptides 84-166 (83 peptides).

The SARS-CoV-2 SNMO defined peptide pool contains 47 synthetic peptides from the human SARS-CoV-2 virus. The peptides are derived from the spike, nucleoprotein, membrane protein, ORF3a and ORF7a.

Recommendation

Product details

Product	FluoroSpot Path: SARS-CoV-2 (S1scan+SNMO) 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. Peptide pools 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 from three COVID-19 convalescent individuals, previously confirmed SARS-CoV-2 positive with PCR, and PBMC from one individual collected before the SARS-CoV-2 pandemic were incubated overnight in the absence or presence of the SARS-CoV-2 S1 scanning pool and anti-CD28 mAb. The number of cells secreting IFN-γ, IL-2 and both analytes was analyzed by FluoroSpot using Mabtech IRIS.

Human PBMC from COVID-19 convalescent individuals (first two from the left from individuals previously confirmed COVID-19 positive with PCR, and third from the left from individual recovering from COVID-19 symptoms) and healthy control. Cells (350,000 cells/well) were incubated overnight in the absence or presence of the SARS-CoV-2 SNMO defined peptide pool in the presence of anti-CD28 mAb. The number of cells secreting IFN-γ and IL-2 was analyzed by FluoroSpot and counted using MabtechIRIS.

Documents

Datasheet/Protocol
SDS

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.

Product highlights

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 June 30, 2020

T cell immunity in COVID-19

Much emphasis has been put on antibody responses, but also T cells are important in the context of COVID-19. In this study, the authors characterized SARS-CoV-2-specific T cell responses by using an extensive FACS panel, peptide stimulations, tetramer staining, and Mabtech IFN-γ ELISpot.

Highlighted research, Published May 30, 2020

B cell FluoroSpot for sensitive ASC detection

Mabtech IRIS and IgG/IgA/IgM FluoroSpot were used to study antibody-secreting plasma cells (ASCs) specific for the nucleocapsid (N) protein. FluoroSpot was shown to be more sensitive than e.g., ELISA.

Highlighted research, Published May 11, 2020

Longevity of immunity against SARS-CoV-2

Investigation of specific T cell responses to SARS-CoV 11 years after outbreak using Mabtech human IFN-γ ELISpot.

Publications (17)

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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