ELISpot Path: SARS-CoV-2 (S+NMO) Human IFN-γ

ELISpot Path: SARS-CoV-2 (S+NMO) Human IFN-γ

3420-4AST-P12-1

Publications: 16
Documents
Questions?
A straightforward setup for functional analysis of a specific scientific question.

Components

Detection mAb7-B6-1, biotin
Enzyme conjugateStreptavidin-ALP
SubstrateBCIP/NBT-plus
PlatePre-coated ELISpot plate, strip (mAb 1-D1K)
Positive controlAnti-CD3 mAb (CD3-2)
Co-stimulusAnti-CD28 mAb (CD28-A)
Peptide poolsSARS-CoV-2 (S), human
SARS-CoV-2 (NMO), human

In stock

Delivery 4-9 business days

Shipping $0

Product specifications

Intended use

This ELISpot Path kit is intended for the enumeration of cells secreting human IFN-γ in response to SARS-CoV-2 (S+NMO) peptides using the ELISpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

Peptide pool composition
The SARS-CoV-2 S defined peptide pool contains 100 peptides from the human SARS-CoV-2 virus. The peptides are derived from the spike (S) protein.
The SARS-CoV-2 NMO defined peptide pool contains 101 peptides from the human SARS-CoV-2 virus. The peptides are derived from the nucleoprotein (N), membrane protein (M), and open reading frame (ORF) proteins (O): ORF1, nsp3, ORF-3a, ORF-7a, and ORF8.
Published HLA types covered by the SARS-CoV-2 NMO defined peptide pool are listed in the datasheets.

Recommendation

Product details

ProductELISpot Path: SARS-CoV-2 (S+NMO) Human IFN-γ
ApplicationELISpot
AnalyteIFN-γ
ReactivityHuman
Specificity

This kit is based on a matched pair of mAbs specific for human IFN-γ.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store antibodies, enzyme conjugate, and substrate at 4-8 °C. Peptide pools should be stored at -20 °C or below. Plates may be kept at room temperature.

Shelf lifeAt least 18 months from date of receipt.
Human PBMC (250,000 cells/well) from one COVID-19 vaccinated individual (mRNA vaccine with spike sequence), one COVID-19 convalescent individual, and from one individual collected before the SARS-CoV-2 pandemic were incubated overnight in the absence or presence of the SARS-CoV-2 S defined peptide pool, SARS-CoV-2 NMO defined peptide pool in the presence of anti-CD28 mAb. The number of cells secreting IFN-γ was analyzed by FluoroSpot using 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 November 7, 2022

7 common ELISpot errors and how to avoid them
ELISpot troubleshooting. We discuss 7 of the most common issues we’ve encountered in ELISpot, what mistakes causes them and how to avoid and correct them.

Tutorial, Published October 14, 2022

What is a positive ELISpot response?
DFR is a non-parametric statistical test for determining positive responses in ELISpot and FluoroSpot.

Tutorial, Published September 2, 2022

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

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.
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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
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FluoroSpot Path: SARS-CoV-2 (S+NMO) Human IFN-γ/IL-2