uPA (urokinase-type plasminogen activator) and tPA (tissue plasminogen activator), which are serine proteases and members of the trypsin family, are essential to the intrinsic coagulation system. tPA is primarily involved in fibrinolysis, whereas uPA principally mediates cell migration and tissue remodeling processes. uPA and tPA are responsible for cleaving plasminogen, a large serum integral-globulin that is deposited on the Fibrin strands within a thrombus. uPA and tPA preferentially target plasminogen at the Arg-Val bond to produce plasmin (also designated fibrinolysin), which is a trypsin-like enzyme that acts on Arg-Lys bonds in Fibrin and Fibrinogen and contributes to the systematic activation of the coagulation cascade. uPA and tPA each consist of two chains that are designated A and B. The A chain of uPA can be cleaved, resulting in low and high molecular mass forms. uPA and tPA are regulated by the serpin family members PAI-1 and PAI-2, which are serine proteinase inhibitors that complex with uPA, tPA and other targeted proteinases and then slowly disassociate to produce cleaved species that fold into stable inactive conformations.
Sample Type:Cell culture supernatants, serum, plasma, and tissue
Intended Use:The Human t-PA BioAssay™ ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human t-PA concentrations in cell culture supernatants, serum, plasma, and tissue.
Sensitivity:7pg/ml
Range:15.625-1000pg/ml
Specificity:Recognizes human t-PA.
Sample Volume:100ul/well
Test Principle:This assay employs the Sandwich immunoassay technique. An anti-h t-PA monoclonal coating antibody is adsorbed onto microwells. t-PA present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-h t-PA antibody is added and binds to t-PA captured by the first antibody. Following incubation unbound Biotinylated anti-h t-PA antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h t-PA antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of t-PA present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven t-PA standard dilutions and t-PA sample concentration determined.
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
