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A miniaturized assay for measuring small molecule phosphorylation in the presence of complex matrices

Spry, Christina; Saliba, Kevin; Strauss, Erick

Description

We describe here a simple, miniaturized radiation-based phosphorylation assay that can be used to monitor phosphorylation of a diverse range of small molecule substrates in the presence of purified and crude enzyme preparations. Ba(OH)2 and ZnSO4 are used to terminate phosphoryl transfer and to precipitate selectively the phosphorylated reaction product in a single step; non-phosphorylated substrate is removed by filtration prior to quantification. The key advantages over alternative...[Show more]

CollectionsANU Research Publications
Date published: 2014
Type: Journal article
URI: http://hdl.handle.net/1885/69921
Source: Analytical Biochemistry
DOI: 10.1016/j.ab.2013.12.010

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