Rat TNF-Alpha ELISA Kit (Tumor Necrosis Factor Alpha)

Full Name: Rat TNF-Alpha ELISA Kit (Tumor Necrosis Factor Alpha)
Reactivity: Rat
Sample Type: Cell Supernatant
Sensitivity: 10.0 pg/ml

BACKGROUND

TNF-alpha is a potent pore-forming molecule that induces an influx of water molecules through ion channels, thereby activating inflammatory reactions. It is also has strong chemotactic effects on neutrophils and lymphocytes, promoting their migration into inflamed tissue. Although it does not possess bacteriocidal activity by itself, it plays a role in stimulating the mechanisms for killing bacteria within macrophages and other phagocytes. The establishment and upkeep of granulomas, which are essential for the defence of intracellular organisms against outside invaders, are mostly dependent on macrophage TNF secretion. Leukocyte trafficking and immune complex clearance are two additional crucial functions of TNF-alpha. It is released by macrophages and cells of the monocyte lineage in response to various stimuli.

TNF-alpha is a protein of 157 amino acids and a molecular weight of 17 kDa. Natural killer cells, T lymphocytes, activated monocytes, and macrophages all make it, and it is primarily found as a homotrimer in various solutions. It is essential for regulating the number of precursor osteoclasts in the bone marrow since doing so can enhance the RANK signalling pathway. TNF-alpha inhibits the recruitment of tyrosine kinase receptors and reduces phosphorylation of CREB and ERK2 in order to attenuate RANK ligand signalling when it binds to its receptor found on osteoclast precursors. It acts on TCR in order to induce signalling cascades that lead to inflammation, autoimmune response and cell death.

INTENDED USE

Rat TNF-alpha ELISA kit is designed for analysing quantitative amounts of rat tumor necrosis factor alpha (TNF-a, TNF-α, TNF-alpha) using cell culture supernatant. This assay has a minimum sensitivity detection limit of 10.0 pg/ml.

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.

Pre-Coated Plate: Anti-rat TNF-α (C) rabbit IgG.
Labelled Antibody (30x Conc.).
Standards (0-8): Recombinant, concentrations 0, 56.25, 112.5, 225, 450, 900, 1,800, 3,600 pg/ml.
EIA Buffer.
Solution For Labelled Antibody.
Chromogen Solution (TMB).
Stop Solution.
Wash Buffer (Concentrated).

SENSITIVITY

The minimum sensitivity detection limit of tumor necrosis factor alpha (TNF-α, TNF-a) using current rat TNF-alpha ELISA kit is 10.0 pg/ml. The dynamic range for this assay is 56.0 – 3,600.0 pg/ml. It has the ability to detect both recombinant and native rat TNF-alpha.

ASSAY CHARACTERISTICS

– Titer Assay: 87 – 102.4%
– Added Recovery Assay: 82.3 – 86.4%
– Intra – Assay: 4.3 – 6.5%
– Inter – Assay: 6.3 – 10.2%
– Specificity (Cross Reactivity): Rat TNF-α (100 %), Rat IL-6 (≦ 0.1 %), Rat MCP-1 (≦ 0.1 %), Rat Osteopontin (≦ 0.1 %), Rat CINC-1 (≦ 0.1 %).
– Sensitivity: 10.55 pg/ml

REFERENCES

The effects of anti-TNF-α antibody on hyper-prolactinemia-related suppression of hCG-induced testosterone release in male rats. J Sex Med. (2012) 9 (4): 1005-14. Huang W.J., et al.
Roles of TNF-α, GSK-3β and RANKL in the occurrence and development of diabetic osteoporosis. Int J Clin Exp Pathol. (2015) 8 (10): 11995-2004. Qi J., et al.
Rat tumour necrosis factor-alpha: expression in recombinant Pichia pastoris, purification, characterization and development of a novel ELISA. Eur Cytokine Netw. (1999) 10 (3): 383-92. Rees G.S., et al.
The role of TNFα in the periaqueductal gray during naloxone-precipitated morphine withdrawal in rats. Neuro-psychopharmacology. (2011) 36 (3): 664-76. Hao S., et al.

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