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ELISpot MAX™ Deluxe Set Human TNF-α

Pricing & Availability
Regulatory Status
RUO
Other Names
Tumor Necrosis Factor alpha, Cachectin, Tumor necrosis factor ligand superfamily member 2
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Product Citations
publications
ELISpot-MAX_Human-TNF-Alpha_121923.png
Human peripheral blood mononuclear cells (PBMC) at 104 cells/well were incubated for 20 hours in the absence or presence of either 100 ng/mL LPS (lipopolysaccharide), 1X Cell Activator without Brefeldin A (Cat. No. 423301), or 1X Cell Activator 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 ELISpot.
  • ELISpot-MAX_Human-TNF-Alpha_121923.png
    Human peripheral blood mononuclear cells (PBMC) at 104 cells/well were incubated for 20 hours in the absence or presence of either 100 ng/mL LPS (lipopolysaccharide), 1X Cell Activator without Brefeldin A (Cat. No. 423301), or 1X Cell Activator 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 ELISpot.
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430224 5 plates £421
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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. It can be expressed and secreted by macrophages, neutrophils, T-cells, NK cells, and a few 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. On the other hand, there is also interest in using TNF-α as an immunostimulant drug to enhance the anti-cancer activity of monoclonal antibodies.

BioLegend’s ELISpot MAX™ Deluxe Set contains pre-optimized essential reagents and additional buffers and solution for Enzyme-Linked Immunosorbent Spot assay. This kit is designed for detection of human TNF-α secreting cells. A monoclonal antibody specific for human TNF-α is coated on ethanol-activated 96-well PVDF membrane plate as the capture. Stimulated and/or unstimulated cells are pipetted directly into the wells, and then incubated at 37oC for a specific period of time. During this period, capture antibody binds to human TNF-α from the secreting cells in the vicinity. Then, cells are removed, and a biotinylated anti-human TNF-α detection antibody is added, followed by avidin-HRP. Subsequently, TMB membrane substrate is added. The TMB membrane substrate reacts with HRP to form a blue-black precipitate at the sites of human TNF-α secreting cells. The end result is visible spots, which corresponds to footprints of TNF-α secreting cells, on the membrane surface.

Product Details
Technical Data Sheet (pdf)

Kit Contents

Kit Contents
  • Human TNF-α ELISpot MAX™ Capture Antibody (200X) 
  • Human TNF-α ELISpot MAX™ Detection Antibody (200X) 
  • Human TNF-α Standard (used as positive control)
  • Avidin-HRP (1000X) 
  • TMB Membrane Substrate
  • Coating Buffer A (5X) 
  • Assay Diluent B (5X)

Product Details

Verified Reactivity
Human
Application

ELISpot
Materials Not Included
  • PVDF-bottom plate: Millipore MultiScreenHTS IP Filter Plate (Cat. No. MSIPS4W10) is recommended. 
  • 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
  • 35% Ethanol
  • 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 
  • Stereomicroscope or an automated ELISpot reader

Antigen Details

Cell Type
Macrophages, Neutrophils, NK cells, T cells
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cancer Biomarkers, Immuno-Oncology, Immunology, Innate Immunity, Neurodegeneration, Signal Transduction
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

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Go To Top Version: 1    Revision Date: 12/19/2023

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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