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  • Title: Time-resolved europium fluorescence in enzyme activity measurements: a sensitive protease assay.
    Author: Karp MT, Suominen AI, Hemmilä I, Mäntsälä PI.
    Journal: J Appl Biochem; 1983 Dec; 5(6):399-403. PubMed ID: 6381465.
    Abstract:
    A method for incorporating into proteins a nonradioactive Eu3+ label, which exhibits fluorescence of a long decay time in the presence of suitable ligands, is described. As an example of the use of this label the method has been developed to work as a sensitive protease assay. By hydrolyzing the Eu3+-labeled casein, bound to an insoluble matrix (Sepharose 4B or Affi-Gel 10), with proteases and measuring the Eu3+ released with a pulsed time-resolved fluorometer it was possible to detect as low as 2.5, 1.0, or 1.0 ng of alpha-chymotrypsin, trypsin, or subtilisin, respectively.
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