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Sphingomyelinase Assays


Sphingomyelinase dose responses were measured in a 96-well white wall/ clear bottom plate with Amplite® Colorimetric Sphingomyelinase Assay Kit using a SpectraMaxR microplate reader (Molecular Devices). As low as 0.08 mU/mL sphingomyelinase was detected with 60 minutes incubation (n=3).
Sphingomyelinase (SMase) is an enzyme that is responsible for cleaving sphingomyelin (SM) to phosphocholine and ceramide. Activation of SMases in cells plays an important role in the cellular responses. Five types of sphingomyelinase have been identified based on their cation dependence and pH optima of action. They are lysosomal acid SMase, secreted zinc-dependent acid SMase, magnesium-dependent neutral SMase, magnesium-independent neutral SMase, and alkaline SMase. Among the five types, the lysosomal acidic SMase and the magnesium-dependent neutral SMase are considered major candidates for the production of ceramide in the cellular response to stress.


Sphingomyelinase dose responses were measured in a 96-well black solid plate with Amplite® Fluorimetric Sphingomyelinase Assay Kit using a Gemini fluorescence microplate reader (Molecular Devices). As low as 0.15 mU/mL sphingomyelinase was detected with 60 minutes incubation (n=3).
Amplite® Colorimetric Sphingomyelinase Assay Kit provides a sensitive method for detecting neutral SMase activity or screening its inhibitors. The kit uses Amplite® Blue as a colorimetric probe to indirectly quantify the phosphocholine produced from the hydrolysis of sphingomyelin by sphingomyelinase (SMase). It can be used for measuring the SMase activity in blood, cell extracts or other solutions. The absorbance of Amplite® Blue at 655 nm is proportional to the formation of phosphocholine, therefore to the SMase activity. The kit is an optimized "mix and read" assay that is compatible with HTS liquid handling instruments.

Amplite® Fluorimetric Sphingomyelinase Assay Kit provides the most sensitive method for detecting neutral SMase activity or screening its inhibitors. The kit uses Amplite® Red as a fluorogenic probe to indirectly quantify the phosphocholine produced from the hydrolysis of sphingomyelin by sphingomyelinase (SMase). It can be used for measuring the SMase activity in blood, cell extracts or other solutions. The fluorescence intensity of Amplite® Red is proportional to the formation of phosphocholine, therefore to the SMase activity. Amplite® Red enables the assay readable either in fluorescence intensity mode or in absorbance mode. The kit is an optimized "mix and read" assay that can be used for real time monitoring of SMase activities.

Amplite® Fluorimetric Acidic Sphingomyelinase Assay Kit provides one of the most sensitive methods for detecting acidic SMase activity or screening its inhibitors. The kit uses Amplite® Red as a fluorogenic probe to indirectly quantify the phosphocholine produced from the hydrolysis of sphingomyelin (SM) by sphingomyelinase (SMase). The fluorescence intensity of Amplite® Red is proportional to the formation of phosphocholine, therefore to the SMase activity. It can be used for measuring the SMase activity in blood, cell extracts or other solutions. The kit is an optimized "mix and read" assay which is compatible with HTS liquid handling instruments.
 

Table 1. Sphingomyelinase Assay Kits

Cat No.
Product Name
Ex (nm)
Em (nm)
Unit Size
13620Amplite® Colorimetric Sphingomyelinase Assay Kit *Blue Color* 655 200 Tests
13622Amplite® Fluorimetric Acidic Sphingomyelinase Assay Kit *Red Fluorescence* 571585200 Tests
13625Amplite® Fluorimetric Sphingomyelin Assay Kit *Red Fluorescence* 571585100 Tests
13621Amplite® Fluorimetric Sphingomyelinase Assay Kit *Red Fluorescence* 571585200 Tests