FluoroSpot Plus: Mouse IFN-γ/IL-10/IL-5

FSP-414743-2

Publications: 1

Documents

Questions?

Ensure reproducibility when diving into a multiplex analysis.

Components

Detection mAbs	Anti-IFN-γ mAb (R4-6A2), BAM
	Anti-IL-10 mAb (51E6), biotin
	Anti-IL-5 mAb (TRFK4), WASP
Fluorophore conjugates	Anti-BAM mAb, 490
	SA-550
	Anti-WASP mAb, 640
Co-stimulus	Anti-CD28 mAb
Buffer/Solution	FluoroSpot enhancer
Plate	Pre-coated FluoroSpot plate (mAbs AN18, MT60, and TRFK5)

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Delivery within 3 weeks

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Product specifications

Intended use

This FluoroSpot Plus kit is intended for the enumeration of cells secreting mouse IFN-γ, IL-10, and IL-5 using the FluoroSpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

Recommendation

Product details

Product	FluoroSpot Plus: Mouse IFN-γ/IL-10/IL-5
Application	FluoroSpot
Analyte	IFN-γ, IL-10, IL-5
Reactivity	Mouse
Specificity	This kit is based on matched pairs of mAbs specific for mouse IFN-γ, IL-10, and IL-5.

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

Mouse splenocytes (100,000 cells/well) were incubated for 48 hours in the absence or presence of ConA. The number of cells secreting IFN-γ, IL-22 and IL-17A 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 (IL-22) and LED640, Cy5 (IL-17A). 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 October 14, 2022

Isolation, freezing, and thawing of splenocytes

This is a protocol on how to best (i) isolate, (ii) freeze, and (iii) thaw splenocytes.

Publications (1)

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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, If2f, Ifg
Cell type	T cell, Tc, Th1, NK cell
Gene ID	15978

IL-10

Analyte description	Interleukin 10 (IL-10) is a pleiotropic cytokine produced by activated T cells and monocytes. It can modulate the functions of immune responses in both a stimulatory and an inhibitory way. IL-10 is thereby involved in the regulation of immune reactions and inflammatory responses.
Alternative names	Interleukin-10, IL-10, IL10, CSIF, If2a, Il-10
Cell type	T cell, Treg, Monocyte/MΦ, mDC
Gene ID	16153

IL-5

Analyte description	Interleukin 5 (IL-5) is a pleiotropic cytokine primarily produced by T and NK cells. IL-5 specifically controls the survival, differentiation, and chemotaxis of eosinophils. But initially, IL-5 was characterized by its ability to support the growth and terminal differentiation of B cells.
Alternative names	Interleukin-5, IL-5, IL5, BCGF-2, Il-5
Cell type	T cell, Th2, NK cell
Gene ID	16191