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Time-Resolved Fluorescence-Based Assay for the Determination of Alkaline Phosphatase Activity and Application to the Screening of Its Inhibitors

Schrenkhammer, Petra ; Rosnizeck, Ina C. ; Dürkop, Axel ; Wolfbeis, Otto S.
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Date of publication of this fulltext: 18 Mar 2011 07:40


Abstract

A single-step end point method is presented for determination of the activity of the enzyme alkaline phosphatase (ALP) using the effect of enhancement of fluorescence of the easily accessible europium(III)-tetracycline 3:1 complex (Eu3TC). Its luminescence, peaking at 616 nm if excited at 405 nm, is enhanced by a factor of 2.5 in the presence of phosphate. Phenyl phosphate was used as a substrate ...

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