Amplified colorimetric assay of alkaline phosphatase using riboflavin 4'-phosphate: a simple method for measuring riboflavin and riboflavin 5'-phosphate.
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.
Harbron
S
S
London Biotechnology Ltd., Department of Biochemistry and Molecular Biology, University College London, England.
Eggelte
H J
HJ
Rabin
B R
BR
D000429:Alcohol Oxidoreductases / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000032:analysis*; D003124:Colorimetry / Q000379:methods*; D005486:Flavin Mononucleotide / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism; D006016:Glycolates / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000032:analysis; D012256:Riboflavin / Q000032:analysis / Q000378:metabolism
D006016:Glycolates; D005486:Flavin Mononucleotide; D006861:Hydrogen Peroxide; D000429:Alcohol Oxidoreductases; C020821:glycollate oxidase; D000469:Alkaline Phosphatase; D012256:Riboflavin