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EK000010-EK0375: Mouse IFNγ ELISA Kit
Range 31.2pg/ml-2000pg/ml
Sensitivity < 5 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of mouse IFNγ in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Interferon-gamma (IFN-gamma) is an inflammatory cytokine that has been implicated in the development of fibrosis in inflamed tissues. The production of IFN-gamma, which is under genetic control, can influence the development of fibrosis in lung allografts.1 IFN-gamma is also produced by natural killer (NK) cells and most prominently by CD8 cytotoxic T cells, and is vital for the control of microbial pathogens.2 Interferon gamma is believed to be crucial for host defence against many infections. Genetically determined variability in IFN-gamma and expression might be important for the development of tuberculosis.3 IFN-gamma activates human macrophage oxidative metabolism and antimicrobial activity.4 In addition to having antiviral activity, IFN-gamma has important immunoregulatory functions. IFN-gamma plays an important role in the control of neointima proliferation.5 The standard product used in this kit is recombinant human IFN?, consisting of 144 amino acid with the molecular mass of 17KDa
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. Awad, M.; Pravica, V.; Perrey, C.; El Gamel, A.; Yonan, N.; Sinnott, P. J.; Hutchinson, I. V. CA repeat allele polymorphism in the first intron of the human interferon-gamma gene is associated with lung allograft fibrosis. Hum. Immunol. 60: 343-346, 1999.
2. Szabo, S. J.; Sullivan, B. M.; Stemmann, C.; Satoskar, A. R.; Sleckman, B. P.; Glimcher, L. H. Distinct effects of T-bet in T(H)1 lineage commitment and IFN-gamma production in CD4 and CD8 T cells. Science 295: 338-342, 2002.
3. Rossouw, M.; Nel, H. J.; Cooke, G. S.; van Helden, P. D.; Hoal, E. G. Association between tuberculosis and a polymorphic NF-kappa-B binding site in the interferon gamma gene. Lancet 361: 1871-1872, 2003.
4. Nathan, C. F.; Murray, H. W.; Wiebe, M. E.; Rubin, B. Y. Identification of interferon-gamma as the lymphokine that activates human macrophage oxidative metabolism and antimicrobial activity. J. Exp. Med. 158: 670-689, 1983.
5. Zohlnhofer, D.; Richter, T.; Neumann, F.-J.; Nuhrenberg, T.; Wessely, R.; Brandl, R.; Murr, A.; Klein, C. A.; Baeuerle, P. A. Transcriptome analysis reveals a role of interferon-gamma in human neointima formation. Molec. Cell 7: 1059-1069, 2001.
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