An automated fluorometric assay for alkaline phosphatase using 3-O-methylfluorescein phosphate.

H D Hill, G K Summer, M D Waters

Index: Anal. Biochem. 24 , 9-17, (1968)

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Abstract

Refrigenation of the substrate 3- O-methylfluoreseein phosphate and attention to cleanliness of the glassware conferred sufficient stability to allow development of an automated fluorometric procedure for measurement of alkaline phosphatase activity at rates of up to 60 samples per hour over the range 0.2–5.6 picomoles/ml of 3- O-methylfluorescein liberated per minute incubation at 37°C. Optimum reagent concentrations were determined which produced linear relationships between fluorescence produced by increasing concentrations of enzyme and by increasing time of incubation.