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Automated measurement of proteinase activity with a fluorogenic substrate using an inverted fluorescence microscope

✍ Scribed by M.H. Dresden; J.P. Rotmans; A.M. Deelder; G. Koper; J.S. Ploem


Publisher
Elsevier Science
Year
1982
Tongue
English
Weight
289 KB
Volume
126
Category
Article
ISSN
0003-2697

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✦ Synopsis


An automated method for measurement of proteinase activities using fluorogenic substrates is described. Enzyme assays were performed' in polystyrene microtitration trays as normally used for the enzyme-linked immunosorbent assay technique. The reaction products were measured using an inverted fluorescence microscope equipped with a photometer. Dam acquisition and processing were via a microcomputer. An acidic thiol-dependent proteinase was used to illustrate the utility of this technique.

' To whom correspondence should be addressed. ' Abbreviation used: ELBA, enzyme-linked immunosorbent assay.

INSTRUMENTATION AND MEASUREMENTS

The optical measurement system utilized for these experiments was essentially as described by Deelder et al. ( 1) for their development of the "Aperture Defined Microvol-