FluoroSpot Plus: Human IFN-γ/Granzyme B/TNF-α

FluoroSpot Plus: Human IFN‑γ/Granzyme B/TNF‑α

FSP-013609-2

Publications: 5

Documents

Questions?

Ensure reproducibility when diving into a multiplex analysis.

Components

Plate	Pre-coated FluoroSpot plate (mAbs 1-D1K, MT34B6, and MT25C5)
Detection mAbs	Anti-IFN-γ mAb (7-B6-1), BAM
	Anti-Granzyme B (MT28), biotin
	Anti-TNF-α mAb (MT20D9), WASP
Fluorophore conjugates	Anti-BAM mAb, 490
	SA-550
	Anti-WASP mAb, 640
Buffer/Solution	FluoroSpot enhancer
Positive control	Anti-CD3 mAb (CD3-2)
Co-stimulus	Anti-CD28 mAb (CD28-A)

In stock

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, granzyme B, and TNF-α 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/Granzyme B/TNF-α
Application	FluoroSpot
Analyte	IFN-γ, Granzyme B, TNF-α
Reactivity	Human
Specificity	This kit is based on matched pairs of mAbs specific for human IFN-γ, granzyme B, and TNF-α.

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 for 42 hours in the absence or presence of a mixture of peptide pools: CEFTA peptide pool for human CD4 T cells (product code 3617-1), CEF extended peptide pool for human CD8 T cells (product code 3618-1), and CMV peptide pool for human CD4 and CD8 T cells (product code 3619-1) and anti-CD28 antibody. The numbers of cells secreting IFN-γ, Granzyme B and TNF-α was analyzed by FluoroSpot. Single cytokine secreting cells as well as double- and triple-secreting cells were determined. Spot center was used to identify spots from double- and triple-secreting cells, visualized in an image overlay of LED490, FITC (IFN-γ) and LED550, Cy3 (Granzyme B) and LED640, Cy5 (TNF-α). Spot analysis was made with 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.

Publications (5)

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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

TNF-α

Analyte description	Tumor necrosis factor (TNF), also known as TNF-α, is produced by many different cell types, e.g., monocytes, macrophages, T cells, and B cells. Among the many effects of TNF-α are protection against bacterial infection, cell growth modulation, immune system regulation, and involvement in septic shock.
Alternative names	Tumor necrosis factor-α, TNF-α, TNF-alpha, TNF-a, TNFa, Tumor necrosis factor-alpha, TNF, DIF-alpha, TNFA, TNFSF2, TNLG1F
Cell type	T cell, Tc, Th1, Th2, Th17, Tfh, Monocyte/MΦ
Gene ID	7124