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FluoroSpot MAX™ Deluxe Set Human TNF-α

Pricing & Availability
Regulatory Status
RUO
Other Names
Tumor Necrosis Factor α (TNF-α), Cachectin, Cytotoxic Factor (CF), Hemorrhagic Factor, Macrophage Cytotoxic Factor (MCF)
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Product Citations
publications
FluoroSpot-MAX_Human-TNF-a_072125
Human PBMC (15,000 cells / well) were incubated for 5 hours in the absence or presence of either 1X Cell Activation Cocktail (without Brefeldin A) (Cat. No. 423301) or 1X Cell Activation Cocktail (with Brefeldin A) (Cat. No. 423303). BioLegend’s Cell Activator cocktail contains PMA and ionomycin at the optimized concentration. Brefeldin A functions to inhibit intracellular protein transport, as a result, it inhibits secretion of cytokines such as TNF-α. The number of cells secreting Human TNF-α was analyzed by FluoroSpot and quantified using an image cytometer equipped with the required fluorescence filter.
  • FluoroSpot-MAX_Human-TNF-a_072125
    Human PBMC (15,000 cells / well) were incubated for 5 hours in the absence or presence of either 1X Cell Activation Cocktail (without Brefeldin A) (Cat. No. 423301) or 1X Cell Activation Cocktail (with Brefeldin A) (Cat. No. 423303). BioLegend’s Cell Activator cocktail contains PMA and ionomycin at the optimized concentration. Brefeldin A functions to inhibit intracellular protein transport, as a result, it inhibits secretion of cytokines such as TNF-α. The number of cells secreting Human TNF-α was analyzed by FluoroSpot and quantified using an image cytometer equipped with the required fluorescence filter.
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430226 5 plates 616€
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Description

Human TNF-α (Tumor Necrosis Factor α), also known as Cachectin, Cytotoxic Factor (CF), is a cytokine which has important roles in regulating a wide range of biological functions from immune activation, cellular differentiation, proliferation, apoptosis, and energy metabolism. TNF-α is expressed and secreted by macrophages, neutrophils, T-cells, NK cells, and various other cell types. TNF-α can bind to two receptors: TNFR1 and TNFR2. Signaling through TNFR1 is mostly pro-inflammatory, while TNFR2 signaling is mainly anti-inflammatory and proliferative. Because of its diverse functions, dysregulation or TNF-α expression and activity has been implicated in many diseases from autoimmune, cancer, to neurodegeneration.

BioLegend’s FluoroSpot MAX™ Deluxe Set contains pre-optimized essential reagents, buffers, and solutions for the detection of human TNF- α secreting cells. In this assay, anti-human TNF- α capture antibodies are coated on a clear-bottom microplate. Cells (stimulated or unstimulated) are added to the wells and incubated, allowing secreted TNF- α to bind to nearby capture antibodies. After incubation, cells are removed, and biotinylated detection antibodies are added followed by Streptavidin-PE. The resulting red spots represent the “footprint” of an individual TNF- α-secreting cell, allowing for precise quantification of cytokine-producing cells in the sample.

Product Details
Technical Data Sheet (pdf)

Kit Details

Kit Contents
  • Human TNF-α Capture Antibody (200X) 
  • Human TNF-α Detection Antibody (200X) 
  • Human TNF-α Standard (used as positive control) 
  • FluoroSpot SA-PE (1000X) 
  • Coating Buffer A (5X) 
  • Assay Diluent B (5X)
Materials Not Included
  • Flat Bottom High Bind Microplate: Corning® Clear Bottom Black Polystyrene Microplates, 96 Well (Catalog No. 3601) is recommended, or equivalent.
  • Black Microplate Lid: Corning® Universal Black Microplate Lid with Corner Notch (Catalog No. 3935) is recommended, or equivalent
  • Wash Buffer: BioLegend Cat. No. 421601 is recommended, or PBS + 0.05% Tween-20  
  • Plate Sealers: BioLegend Cat. No. 423601 is recommended 
  • 0.22 μm filters
  • Sterile PBS (Phosphate-Buffered Saline): 8.0 g NaCl, 1.16 g Na2HPO4, 0.2 g KH2PO4, 0.2 g KCl, add deionized water to 1 L; pH to 7.4, 0.22 μm filtered
  • Sterile cell culture media
  • 37oC CO2 incubator
  • Deionized (DI) water 
  • Adjustable pipettes to measure volumes ranging from 2 μL to 1 mL 
  • Wash bottle, manifold dispenser, or automated microplate washer 
  • Tubes for dilution
  • Timer
  • Alcohol Wipes 
  • Revvity® CeligoTM Image Cytometer (Catalog No. 200-BFFL-5C) is recommended , or equivalent image cytometer with Excitation 531/40 nm and Emission 629/53 nm.
  • Optional: Carboxyl Beads B4 (Cat. No. 740175)

Product Details

Verified Reactivity
Human
Application

FluoroSpot

Antigen Details

Cell Type
B cells, Eosinophils, Macrophages, Mast cells, Neutrophils, T cells
Biology Area
Cancer Biomarkers, Immuno-Oncology, Immunology, Innate Immunity
Molecular Family
Cytokines/Chemokines, Innate Immune Signaling
Gene ID
7124 View all products for this Gene ID
UniProt
View information about TNF-alpha on UniProt.org

Related FAQs

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Go To Top Version: 1    Revision Date: 04/22/2025

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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