title: A simple spectrophotometric assay of plasminogen activator: comparison with the fibrinolytic method.
abstract: We describe a simple assay of plasminogen activator in which the enzyme reacts with a mixture of plasminogen and H-D-valyl-L-leucyl-L-lysine-p-nitroanilide for 1 h at 37 degrees C after which the absorbance is measured at 405 nm. The method detects as little as 2 CTA milliunits of activator and is linear over a 100-fold range of enzyme concentration. The new procedure has been used successfully for the assay of activator in breast tumor cytosols, cell culture supernatants, and pleural or ascitic fluids. Thirty-two biological samples have been assayed for plasminogen activator activity with both the spectrophotometric method and a classical fibrinolytic technique using radiolabeled fibrin. Although the two series of results are significantly correlated, the activities measured with the former assay are significantly different from those determined with the latter. It is shown that the spectrophotometric method is, in many respects, superior to the fibrinolytic procedure.
mesh_terms: D001202:Ascitic Fluid / Q000378:metabolism; D001943:Breast Neoplasms / Q000032:analysis; D002478:Cells, Cultured; D003600:Cytosol / Q000032:analysis; D005260:Female; D005342:Fibrinolysis; D006801:Humans; D010960:Plasminogen Activators / Q000032:analysis*; D010996:Pleural Effusion / Q000378:metabolism; D013053:Spectrophotometry / Q000379:methods
chemical_list: D010960:Plasminogen Activators