Description
Principle of the Assay: This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-uPA polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-uPA polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the uPA amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of uPA can be calculated.
Background: Urokinase (trade name Abbokinase), also called urokinase-type plasminogen activator (uPA), is a 411-residue protein, consisting of three domains: the serine protease domain, the kringle domain, and the growth factor domain. It 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.[1] Activation of plasmin triggers a proteolysis cascade that, depending on the physiological environment, participates in thrombolysis or extracellular matrix degradation. This links urokinase to vascular diseases and cancer