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EK000044-EK0535: Human uPA ELISA Kit
Range 62.5pg/ml-4000pg/ml
Sensitivity < 5 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitative detection of human uPA in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Plasminogen activator, urokinase (PLAU, uPA) converts plasminogen to plasmin. Plasmin is involved in processing of amyloid precursor protein and degrades secreted and aggregated amyloid-beta, a hallmark of Alzheimer disease (AD).1 Urokinase has a molecular mass of about 54 kD and is composed of 2 disulfide-linked chains, A and B, of molecular masses 18 kD and 33 kD, respectively. It localized on 10q24. uPA facilitates cell migration by localizing proteolisys on the cell surface and by inducing intracellular signalling pathways. In human vascular smooth muscle cell (VSMC), uPA stimulates migration via the uPA receptor (uPAR) signalling complex containing TYK2 and phosphatidylinositol 3-kinase (PI3-K).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. Finckh, U.; van Hadeln, K.; Muller-Thomsen, T.; Alberici, A.; Binetti, G.; Hock, C.; Nitsch, R. M.; Stoppe, G.; Reiss, J.; Gal, A. : Association of late-onset Alzheimer disease with a genotype of PLAU, the gene encoding urokinase-type plasminogen activator on chromosome 10q22.2. Neurogenetics 4: 213-217, 2003.
2. Kiian, I.; Tkachuk, N.; Haller, H.; Dumler, I. : Urokinase-induced migration of human vascular smooth muscle cells requires coupling of the small GTPase RhoA and Rac1 to the Tyk2/PI3-K signalling pathway. Thromb. Haemost. 89: 904-914, 2003.
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