The Neutral Sphingomyelinase Assay Service provides a sensitive and homogenous method to measure the activity of sphingomyelinase in vitro. In this enzyme coupled assay sphingomyelinase catalyzes the hydrolysis of sphingomyelin into phosphorylcholine and ceramide. Alkaline phosphatase catalyzes the hydrolysis of phosphorylcholine and the oxidation of choline to hydrogen peroxide is catalyzed by choline oxidase. DAOS plus 4-Aminoantipyrine (4 AAP) in the presence of hydrogen peroxide and peroxidase results in the oxidative coupling of DAOS and 4 AAP to form a blue chromogen that is detected by measuring the absorbance of light at 595 nm. Experiments have shown the Neutral Sphingomyelinase Assay Service measures the activity of Sphingomyelinase at concentrations as small as 78 μU/mL.
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