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EK000096-EK0536: Human uPAR ELISA Kit
Range 62.5pg/ml-4000pg/ml
Sensitivity < 4 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of human uPAR in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
The urokinase-type plasminogen activator receptor (uPAR) is a key molecule in the regulation of cell-surface plasminogen activation and, as such, plays an important role in many normal as well as pathological processes.1 The cDNA for Mo3, an activation antigen expressed by human monocytes and myelomonocytic cell lines after stimulation by a variety of agents. Mo3 expression in vivo is associated predominantly with macrophages in inflammatory sites. It is a highly glycosylated protein of about 50 kD in monocytes where it is anchored to the plasma membrane by glycosyl-phosphatidylinositol linkage. The complete coding sequence of the cDNA has been found to encode 335 amino acids including a predicted signal peptide of 22 residues and a hydrophobic C-terminal portion. Mo3 is identical to the human receptor for the urokinase plasminogen activator.2 UPAR is a useful prognostic marker for biologically aggressive forms of endometrial cancer.3 PLAUR is located at chromosome 19q13.1-q13.2.1 The standard product used in this kit is recombinant human uPAR, consisting of 287 amino acids with the molecular mass of 31KDa.
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. Borglum, A. D.; Byskov, A.; Ragno, P.; Roldan, A. L.; Tripputi, P.; Cassani, G.; Dano, K.; Blasi, F.; Bolund, L.; Kruse, T. A. Assignment of the urokinase-type plasminogen activator receptor gene (PLAUR) to chromosome 19q13.1-q13.2. Am. J. Hum. Genet. 50: 492-497, 1992.
2. Min, H. Y.; Semnani, R.; Mizukami, I. F.; Watt, K.; Todd, R. F., III; Liu, D. Y. cDNA for Mo3, a monocyte activation antigen, encodes the human receptor for urokinase plasminogen activator. J. Immun. 148: 3636-3642, 1992.
3. Memarzadeh, S.; Kozak, K. R.; Chang, L.; Natarajan, S.; Shintaku, P.; Reddy, S. T.; Farias-Eisner, R. Urokinase plasminogen activator receptor: prognostic biomarker for endometrial cancer. Proc. Nat. Acad. Sci. 99: 10647-10652, 2002. Note: Erratum: Proc. Nat. Acad. Sci. 99: 12501 only, 2002.
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