Amplified luminometric assays of alkaline phosphatase using riboflavin phosphates.
An assay for alkaline phosphatase is described which is based on the hydrolysis of riboflavin phosphates (5'FMN or 4'FMN) to produce riboflavin. This is converted to 5'FMN using riboflavin kinase, and then assayed using the bacterial bioluminescent system from Vibrio harveyi or V. fischeri. The most sensitive assay is obtained using 4'FMN, which can measure less than 20 amol after a 1-hour incubation.
Harbron
S
S
London Biotechnology Ltd, Department of Biochemistry and Molecular Biology, University College London, UK.
Eggelte
H J
HJ
Benson
S M
SM
Rabin
B R
BR
D000469:Alkaline Phosphatase / Q000032:analysis*; D038181:FMN Reductase; D005486:Flavin Mononucleotide / Q000378:metabolism*; D007700:Kinetics; D008156:Luciferases / Q000302:isolation & purification / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism; D010770:Phosphotransferases / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor); D014733:Vibrio / Q000201:enzymology
D008164:Luminescent Proteins; D005486:Flavin Mononucleotide; D008156:Luciferases; D038181:FMN Reductase; D009247:NADH, NADPH Oxidoreductases; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C022532:riboflavin kinase; D000469:Alkaline Phosphatase