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A fluorescence-based assay for 2-oxoglutarate-dependent oxygenases.

Abstract:

A widely used generic assay for 2-oxoglutarate-dependent oxygenases relies upon monitoring the release of 14CO2 from labeled [1-14C]-2-oxoglutarate. We report an alternative assay in which depletion of 2-oxoglutarate is monitored by its postincubation derivatization with o-phenylenediamine to form a product amenable to fluorescence analysis. The utility of the procedure is demonstrated by assays with hypoxia-inducible factor hydroxylases where it was shown to give results similar to those rep...

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Publication status:
Published

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Publisher copy:
10.1016/j.ab.2004.09.019

Authors


McNeill, LA More by this author
Hewitson, KS More by this author
More by this author
Institution:
University of Oxford
Department:
Oxford, MPLS, Chemistry, Organic Chemistry
Journal:
Analytical biochemistry
Volume:
336
Issue:
1
Pages:
125-131
Publication date:
2005-01-05
DOI:
EISSN:
1096-0309
ISSN:
0003-2697
URN:
uuid:e57884e0-2b9b-4051-baa6-8e22495d0988
Source identifiers:
32927
Local pid:
pubs:32927

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