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A simple, sensitive and specific assay for plasminogen activators is described. The assay utilizes fluorescein-labeled fibrinogen or fibrin at low concentrations, and enables simultaneous evaluation of the plasminogen and fibrin dependence of the reaction, that is, discrimination of tissue-type and urokinase-type plasminogen activators, and non-specific proteolysis. Addition of antisera verify identification of the activator species. The assay reagent contains plasminogen and fluorescein-labeled fibrinogen, to which is added the specimen and then then thrombin, either at the initiation or the termination of the reaction. Supernatant fluorescence is proportional to plasminogen activator concentration. With a four-hour incubation, 1 milliunit (14 pg) of tissue (melanoma) plasminogen activator (TPA) or 2 milliunit (36 pg) of urokinase (UK) may be detected.
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Accepted: October 9, 1984
Received: June 22, 1984
© 1985 Published by Elsevier Inc.