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EK000047-EK0584: Human TNFβ ELISA Kit
Range 62.5pg/ml-4000pg/ml
Sensitivity < 5 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of human TNFβ in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Tumor necrosis factor-beta (TNF-beta), previously called lymphotoxin-alpha (LTA), is a cytokine produced by lymphocytes.1 TNF-alpha and TNF-beta share 35% identity and 50% homology in the amino acid sequence.2 The substance LTA mediates a wide variety of inflammatory, immunostimulatory, and antiviral responses.1 LTA is associated with susceptibility to myocardial infarction, asthma and other diseases.3 The LTA gene is located on human chromosome 6.4 The standard product used in this kit is recombinant protein, consisting of 172 amino acids with the molecular mass of 18.8KDa.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Nakayama, T.; Soma M.; Sato, N.; Haketa, A.; Kosuge, K.; Aoi, N.; Sato, M.; Izumi, Y.; Matsumoto, K.; Kanmatsuse, K.; Kokubun, S. An association study in essential hypertension using functional polymorphisms in lymphotoxin-alpha gene. Am J Hypertens. 2004 Nov;17(11 Pt 1):1045-9.
2. Aggarwal, B. B.; Eessalu, T. E.; Hass, P. E. Characterization of receptors for human tumour necrosis factor and their regulation by gamma-interferon. Nature 318: 665-667, 1985.
3. Knight, J. C.; Keating, B. J.; Kwiatkowski, D. P. Allele-specific repression of lymphotoxin-alpha by activated B cell factor-1. Nature Genet. 36: 394-399, 2004.
4. Nedwin, GE.; Jarrett-Nedwin, J.; Smith, DH.; Naylor, SL.; Sakaguchi, AY.; Goeddel, DV.; Gray, PW. Structure and chromosomal localization of the human lymphotoxin gene. J Cell Biochem. 1985;29(3):171-81.
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