TNF Alpha ELISA Kit (Tumor Necrosis Factor Alpha)

INTENDED USE

Human TNF alpha ELISA kit is a reliable quantitative procedure for measuring tumor necrosis factor alpha (TNF alpha, TNF-a, TNF-α, TNF-alpha) in human serum, plasma (EDTA, heparin, citrate) and cell culture medium. This assay has a minimum sensitivity detection limit of 0.7 pg/ml.

BACKGROUND

Tumor necrosis factor alpha (TNF alpha) is structurally a homotrimer protein of 157 amino acids that is primarily produced by; natural killer cells, T-lymphocytes and activated macrophages. It can activate a many inflammatory chemicals. It is a soluble and transmembrane form. Transmembrane TNF (tmTNF) is the originally synthesised precursor form and must be converted into soluble TNF (sTNF) by TNF-converting enzyme (TACE), a membrane-bound disintegrin metalloproteinase. It generally sends chemical signals for biological processes like inflammation and cell death by binding to its receptors, primarily TNFR1 and TNFR2. The death domain (DD) that TNFR1 processes interacts with the TNFR1-associated death domain (TRADD) adaptor protein. TNFR1 is activated by both sTNF and tmTNF.

In 1985, the human TNF alpha gene was cloned. It measures around 3 kilobases in length, has 4 exons, and corresponds to chromosome 6p21.3. The last exon is related to lymphotoxin alpha (LTA). An AU-rich fragment can be found in the three primary untranslated region (3′-UTR).

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• Microplate: Anti human TNF-alpha antigen coated microplate wells.
• Conjugate HRP.
• Zero Calibrator.
• Calibrator 1-5: Concentrations of the standard range from 0-518 pg/ml.
• Conjugate Buffer.
• Incubation Buffer.
• Wash Solution (200x Concentrated).
• Controls 1+2 (Lyophilised).
• Chromogen: Tetramethylbenzydine (TMB).
• Stopping Solution: HCl (1.0N).

SENSITIVITY

The minimum sensitivity detection limit of tumor necrosis factor alpha (TNF alpha, TNF-alpha, TNF-α, TNF-a) using current TNF alpha ELISA kit was approximately 0.7 pg/ml. The dynamic range for this assay is 5.35 – 530.0 pg/ml.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for human natural and recombinant TNF-α. No cross reactivity with 50 ng of IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-6, IL-7, IL-8, IL-10, TNF-β, IFN-α, IFN-β, IFN-γ, TGF-β, GM-CSF, OSM , MIP-1α, MIP-1β, LIF, MCP-1, G-CSF and RANTES.
– Intra-Assay Precision: 6.3 – 6.6%
– Inter-Assay Precision: 3.3 – 4.5%
– Recovery (Accuracy): 84 – 108%

REFERENCES

1. Tumor necrosis factor type alpha stimulates human endothelial cells to produce granulocyte/macrophage colony-stimulating factor. Proc. Nat. Acad. Sci. (1986) 83: 7467-7471. Broudy, V.C., et al.
2. Tumor necrosis factor α signaling in macrophages. Crit Rev Eukaryot Gene Expr. (2010) 20 (2): 87-103. Review. Parameswaran N. and Patial S.
3. TNF-alpha and obesity. Curr Dir Autoimmun. (2010) 11: 145-56. Review. Tzanavari T., et al.

ADDITIONAL INFORMATION

• Full Name: TNF Alpha ELISA Kit (Tumor Necrosis Factor Alpha)
• Reactivity: Human
• Sample Type: Serum, Plasma (EDTA, Citrate, Heparin), Cell Culture
• Sensitivity: 0.7 pg/ml

OTHER RELATED ELISA KITS

• IFN Gamma ELISA Kit
• Adiponectin ELISA Kit
• Insulin ELISA Kit