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Amplified colorimetric assay of alkaline phosphatase using riboflavin 4′-phosphate: A simple method for measuring riboflavin and riboflavin 5′-phosphate

✍ Scribed by Stuart Harbron; Henny J. Eggelte; Brian R. Rabin


Publisher
Elsevier Science
Year
1991
Tongue
English
Weight
504 KB
Volume
198
Category
Article
ISSN
0003-2697

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✦ Synopsis


Alkaline phosphatase hydrolyzes riboflavin 4'-phosphate to produce riboflavin. This is converted to riboflavin 5'-phosphate, using riboflavin kinase, which reconstitutes apoglycolate oxidase to give hologlycolate oxidase. This enzyme catalyzes the oxidation of glycolate with simultaneous production of hydrogen peroxide which is detected via the formation of a colored product through the action of peroxidase. The system allows the detection of 4 amol after a 2-h incubation.