The Neutral Sphingomyelinase Activity Assay kit is an enzyme coupled assay that measures sphingomyelinase activity in biological samples through the downstream production of choline.
Assay Incubation time: 2-4 hours
Assay Range: 5 mU/mL – 0.078 mU/mL
Sample Type: tissue homogenates, cell lysates, serum, saliva, and urine
The Neutral Sphingomyelinase Activity Assay kit can detect sphingomyelinase at concentrations as small as 78 µU/mL. The kit provides all reagents to measure the Neutral Sphingomyelinase (nSMase) activity of 40 samples ran in duplicate.
In this enzyme coupled assay, N-SMase catalyzes the hydrolysis of sphingomyelin into phosphorylcholine and ceramide. Alkaline phosphatase then catalyzes phosphorylcholine to choline. Choline is them oxidized by Choline oxidase to form hydrogen peroxide. Finally, 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.