TheNeutralSphingomyelinaseAssayServiceprovidesasensitiveandhomogenousmethodtomeasuretheactivityofsphingomyelinaseinvitro.Inthisenzymecoupledassaysphingomyelinasecatalyzesthehydrolysisofsphingomyelinintophosphorylcholineandceramide.Alkalinephosphatasecatalyzesthehydrolysisofphosphorylcholineandtheoxidationofcholinetohydrogenperoxideiscatalyzedbycholineoxidase.DAOSplus4-Aminoantipyrine(4AAP)inthepresenceofhydrogenperoxideandperoxidaseresultsintheoxidativecouplingofDAOSand4AAPtoformabluechromogenthatisdetectedbymeasuringtheabsorbanceoflightat595nm.ExperimentshaveshowntheNeutralSphingomyelinaseAssayServicemeasurestheactivityofSphingomyelinaseatconcentrationsassmallas78μU/mL.
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