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Updated: Aug 22, 2025

Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples
Published on: May 10, 2015
Fluorometric Characterization of DUB Activity: From Single-Enzyme Reactions to Live Intact Cells
Alireza Rahnama1, Adam T Melvin2
1Cain Department of Chemical Engineering, Louisiana State University, Baton Rouge, LA, USA.
Abstract:
Fluorescently tagged molecular probes capable of time- and concentration-dependent quantification of deubiquitinating enzyme (DUB) activity allow for precise characterization of both enzyme and DUB inhibitor. These probes are compatible with most plate readers allowing for rapid, facile fluorometric analysis of DUB activity. DUB activity can be measured in purified enzyme reactions, in cell lysates, or in intact cells depending upon the choice of the fluorometric probe. This chapter describes protocols and potential analysis tools to investigate DUB activity in these three scenarios.
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