Invitrogen™ Amplex™ Red Sphingomyelinase Assay Kit

Description

• The Amplex Red Sphingomyelinase Assay Kit provides a sensitive, rapid, and simple fluorometric method for detecting very low concentrations of sphingomyelinase using a fluorescence microplate reader or fluorometer
• In this enzyme-coupled assay, sphingomyelinase activity is monitored indirectly using 10-acetyl-3,7-dihydroxyphenoxazine (Amplex Red reagent), a sensitive fluorogenic probe for hydrogen peroxide
• Sphingomyelinase hydrolyses the sphingomyelin to yield ceramide and phosphorylcholine
• Alkaline phosphatase is added, which hydrolyzes phosphorylcholine to form choline
• The choline is then oxidized by choline oxidase to form betaine and hydrogen peroxide
• In the presence of horseradish peroxidase, the hydrogen peroxide reacts with Amplex Red reagent in a 1:1 stoichiometric ratio to generate the highly fluorescent product resorufin
• Because resorufin has absorption and fluorescence emission maxima of approximately 571nm and 585nm, respectively, there is little interference from autofluorescence in most biological samples
• Detects sphingomyelinase activity levels as low as 80μU/mL in a 200μL assay volume Format allows for multiple time point measurements Designed for minimal autofluorescence interference Custom assay design and packaging are also available Use Amplex Red Assays for a Broad Range of Investigations A wide variety of validated Amplex Red assays are available for studying cell signaling and lipids, neurobiology, inflammation and immune function, and metabolism
• We also offer Amplex UltraRed Reagent, a second-generation reagent providing greater sensitivity and brighter fluorescence, and the Amplex Red/UltraRed Stop Reagent
• The Amplex Red/UltraRed Stop Reagent provides convenience and control by allowing the fluorescence signal-generating reaction to be terminated at a user-determined time point
• After addition of the stop reagent, the fluorescence signal remains stable for at least three hours
• Enzyme and Protein Activity Assays, Lipase and Phopholipase Activity, Proteins, Expression, Isolation and Analysis Order Info Shipping Condition: Wet ice