Description
Key facts
Detection method:Colorimetric,
Sample types:Cell culture extracts, Whole Blood,
Assay type:Enzyme activity,
Sensitivity:= 0.08 mU/mL,
Assay Platform:Microplate reader
Product details:
Sphingomyelinase Assay Kit (Colorimetric) (ab138876) provides a sensitive method for detecting neutral SMase activity or screening its inhibitors. The kit uses our proprietary AbBlue Indicator as a colorimetric probe to indirectly quantify the phosphocholine produced from the hydrolysis of sphingomyelin (SM) by sphingomyelinase (SMase).
The sphingomyelinase assay can be used for measuring the SMase activity in blood, cell extracts or other solutions. The absorbance of light 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.
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 (SMase) 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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Acid sphingomyelinase (ASMase) also known as sphingomyelin phosphodiesterase 1 or NP is an enzyme involved in sphingolipid metabolism. ASMase has a mass of approximately 75 kDa and appears in lysosomes where it converts sphingomyelin to ceramide and phosphorylcholine. This enzyme is important in maintaining cellular lipid balance and signaling. Expression of ASMase occurs in various tissues such as the liver spleen and brain.
Biological function summary
ASMase plays a role in membrane microdomain composition through its involvement in ceramide production. It participates in generating ceramide-enriched platforms that facilitate the clustering of signaling molecules. Ceramide acts as a second messenger in multiple cellular processes including apoptosis proliferation and inflammation. ASMase operates in the lysosomal lipid degradation pathway and connects with other lysosomal enzymes to modulate lipid turnovers such as glucosylceramidase affecting downstream cellular functions.
Pathways
Sphingolipid metabolism involves ASMase. This enzyme participates in the ceramide signaling pathway influencing apoptosis and stress responses. Related proteins in this pathway include casein kinase II which phosphorylates ASMase and cathepsin D involved in the lysosomal degradation process. ASMase activity alters ceramide levels impacting pro-apoptotic and pro-survival signals mediated by related proteins in the cell signaling network.
ASMase deficiency connects to Niemann-Pick disease types A and B characterized by lipid accumulation in lysosomes. Mutations in the ASMase gene lead to impaired enzyme function resulting in excessive sphingomyelin storage and cell damage. The disorder links ASMase to proteins such as hexa-beta-N-acetylglucosaminidase which is affected in other lysosomal storage disorders. Research shows that ASMase activity also influences cardiovascular diseases by regulating ceramide and cholesterol levels in atherosclerotic lesions connecting it to inflammatory pathways involving adhesion molecules and cytokines.
