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EK000050-EK0495: Mouse Rantes ELISA Kit
Range 15.6 pg/ml - 1000 pg/ml
Sensitivity < 1 pg/ml
Specificity: No detectable cross-reactivity with any other cytokine.
Application: For quantitative detection of mouse Rantes in sera, plasma, body fluids, tissue lysates or cell culture supernatant.
Expiration: four months at 4°C and eight months at -20°C.
Background
Rantes (regulated on activation normal T cell expressed and secreted) is one of the natural ligands for the chemokine receptor CCR5 and potently suppresses in vitro replication of the R5 strains of HIV-1, which use CCR5 as a coreceptor. Peripheral blood mononuclear cells or CD4(+) lymphocytes obtained from different individuals have wide variations in their ability to secrete RANTES.1 The Rantes gene product is predicted to be 10 kDa and, after cleavage of the signal peptide, approximately 8 kDa. Of the 68 residues, 4 are cysteines, and there are no sites for N-linked glycosylation. Rantes is expressed by cultured T cell lines that are Ag specific and growth factor dependent.2
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. Liu, H.; Chao, D.; Nakayama, E. E.; Taguchi, H.; Goto, M.; Xin, X.; Takamatsu, J.; Saito, H.; Ishikawa, Y.; Akaza, T.; Juji, T.; Takebe, Y.; and 9 others. Polymorphism in RANTES chemokine promoter affects HIV-1 disease progression. Proc. Nat. Acad. Sci. 96: 4581-4585, 1999.
2. Schall, T. J.; Jongstra, J.; Dyer, B. J.; Jorgensen, J.; Clayberger, C.; Davis, M. M.; Krensky, A. M. A human T cell-specific molecule is a member of a new gene family. J. Immun. 141: 1018-1025, 1988.
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