- 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 ELISpot-MAX_Human-TNF-Alpha_121923.png](https://d1spbj2x7qk4bg.cloudfront.net/Admin/Public/GetImage.ashx?Image=/Files/Images/media_assets/products/product_images/ELISpot-MAX_Human-TNF-Alpha_121923.png&Width=240&Height=300&altFmImage_path=&Compression=90&Crop=5)
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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.
Cat # | Size | Price | Quantity Check Availability | Save | ||
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430224 | 5 plates | 615 CHF |
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.
Kit Contents
- Kit Contents
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- 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
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- 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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