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A continuous spectrophotometric assay for alkaline phosphatase with glycerophosphate as substrate
A Bianchi1, E Giachetti, P Vanni
1Dipartimento di Scienze Biochimiche, Università di Firenze, Florence, Italy.
Journal of Biochemical and Biophysical Methods
|January 1, 1994
Abstract:
We describe a continuous coupled spectrophotometric assay for alkaline phosphatase which uses alpha- or beta-glycerophosphate as substrate, and glycerol dehydrogenase as ancillary enzyme. The glycerol liberated by alkaline phosphatase is determined by measuring the increase in absorbance at 340 nm caused by NADH formation that is combined with glycerol oxidation by the ancillary enzyme. The assay procedure was optimized using a bovine bone extract as alkaline phosphatase source.