FluoroSpot Plus: Human IL-1β/IL-6/TNF-α

FluoroSpot Plus: Human IL‑1β/IL‑6/TNF‑α

FSP-121309-2

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Ensure reproducibility when diving into a multiplex analysis.

Components

Plate	Pre-coated FluoroSpot plate (mAbs MT175, 13A5, and MT25C5)
Detection mAbs	Anti-IL-1β mAb (7P10), BAM
	Anti-IL-6 (39C3), biotin
	Anti-TNF-α mAb (MT20D9), WASP
Fluorophore conjugates	Anti-BAM mAb, 490
	SA-550
	Anti-WASP mAb, 640
Buffer/Solution	FluoroSpot enhancer

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Product specifications

Intended use

This FluoroSpot Plus kit is intended for the enumeration of cells secreting human IL-1β, IL-6, 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 IL-1β/IL-6/TNF-α
Application	FluoroSpot
Analyte	IL-1β, IL-6, TNF-α
Reactivity	Human
Specificity	This kit is based on matched pairs of mAbs specific for human IL-1β, IL-6, 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 (5000 cells/well) were incubated for 20 hours in the absence or presence of LPS and the number of cells secreting IL-1β, IL-6 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 (IL-1β) and LED550, Cy3 (IL-6) and LED640, Cy5 (TNF-α). Spot analysis was made with Mabtech IRIS.

Documents

Datasheet/Protocol
SDS

Tutorials

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Tutorial, Published September 2, 2022

Isolation, freezing, and thawing of PBMCs

This is a protocol on how to best isolate, freeze, and thaw PBMCs.

Publications (0)

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Analyte information

IL-1β

Analyte description	Interleukin 1ß (IL-1ß) is a proinflammatory cytokine and inducer of acute phase responses. IL-1ß is produced primarily by monocytes, macrophages, and dendritic cells after induction by microbes.
Alternative names	Interleukin-1ß, IL-1ß, IL-1F2, Interleukin-1beta, IL-1 beta, IL1b, Interleukin-1 beta, IL-1, IL1-BETA, IL1F2, IL1beta
Cell type	Monocyte/MΦ, mDC
Gene ID	3553

IL-6

Analyte description	Interleukin 6 (IL-6) is a pleiotropic cytokine produced by many different cell types and plays a role in a wide range of functions, such as immune responses, acute-phase reactions, and hematopoiesis. Among other things, it augments antibody production from activated B cells in vitro.
Alternative names	Interleukin 6, IL-6, IL6, IFB-B502, BSF-2, BCDF, BSF2, CDF, HGF, HSF, IFN-beta-2, IFNB2
Cell type	B cell, Monocyte/MΦ, mDC
Gene ID	3569

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