FluoroSpot Path: SARS-CoV-2 (S1scan+SNMO) Human IFN-γ/Granzyme B

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

FSP-0136-P1-1

Publications: 0

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Questions?

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

Components

Plate	Pre-coated FluoroSpot plate (mAbs 1-D1K and MT34B6)
Detection mAbs	Anti-IFN-γ mAb (7-B6-1), BAM
	Anti-Granzyme B (MT28), 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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Delivery 4-9 business days

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Product specifications

Intended use

This FluoroSpot Path kit is intended for the enumeration of cells secreting human IFN-γ and granzyme B 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/Granzyme B
Application	FluoroSpot
Analyte	IFN-γ, Granzyme B
Reactivity	Human
Specificity	This kit is based on matched pairs of mAbs specific for human IFN-γ and granzyme B.

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

PBMC from a COVID-19 convalescent individual and PBMC from a healthy control were incubated 48 h in the absence or presence of the SARS-CoV-2 S1 scanning pool and SARS-CoV-2 S N M O defined peptide and anti-CD28 mAb. As a positive control, cells were stimulated with PHA + anti-C28 mAb. The number of cells secreting IFN-γ, Granzyme B and both analytes was analyzed by FluoroSpot using Mabtech IRIS.

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 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 (0)

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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

Granzyme B

Analyte description	Granzyme B is a serine protease released by the cytoplasmic granules within cytotoxic T cells and NK cells. Granzyme B will act together with perforin and other cytotoxic mediators to induce target cell death by apoptosis.
Alternative names	Granzyme B, GzmB, GrB, GrzB, C11, CCPI, CGL-1, CGL1, CSP-B, CSPB, CTLA1, CTSGL1, HLP, SECT
Cell type	T cell, Tc, Th1, NK cell
Gene ID	3002