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Sphingomyelinase Assay Kit (Colorimetric) (ab138876) provides a sensitive method for detecting neutral SMase activity or screening its inhibitors.

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Functional Studies - Sphingomyelinase Assay Kit (Colorimetric) (AB138876), expandable thumbnail

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Key facts

Detection method
Colorimetric
Sample types
Cell culture extracts, Whole Blood
Assay type
Enzyme activity
Reactive species
Mammals
Sensitivity
= 0.08 mU/mL

Target data

Function

Converts sphingomyelin to ceramide (PubMed:12563314, PubMed:1840600, PubMed:18815062, PubMed:25339683, PubMed:25920558, PubMed:27659707, PubMed:33163980). Exists as two enzymatic forms that arise from alternative trafficking of a single protein precursor, one that is targeted to the endolysosomal compartment, whereas the other is released extracellularly (PubMed:20807762, PubMed:21098024, PubMed:9660788). However, in response to various forms of stress, lysosomal exocytosis may represent a major source of the secretory form (PubMed:12563314, PubMed:20530211, PubMed:20807762, PubMed:22573858, PubMed:9393854). In the lysosomes, converts sphingomyelin to ceramide (PubMed:20807762, PubMed:21098024). Plays an important role in the export of cholesterol from the intraendolysosomal membranes (PubMed:25339683). Also has phospholipase C activities toward 1,2-diacylglycerolphosphocholine and 1,2-diacylglycerolphosphoglycerol (PubMed:25339683). Modulates stress-induced apoptosis through the production of ceramide (PubMed:8706124). When secreted, modulates cell signaling with its ability to reorganize the plasma membrane by converting sphingomyelin to ceramide (PubMed:12563314, PubMed:17303575, PubMed:20807762). Secreted form is increased in response to stress and inflammatory mediators such as IL1B, IFNG or TNF as well as upon infection with bacteria and viruses (PubMed:12563314, PubMed:20807762, PubMed:9393854). Produces the release of ceramide in the outer leaflet of the plasma membrane playing a central role in host defense (PubMed:12563314, PubMed:20807762, PubMed:9393854). Ceramide reorganizes these rafts into larger signaling platforms that are required to internalize P. aeruginosa, induce apoptosis and regulate the cytokine response in infected cells (PubMed:12563314). In wounded cells, the lysosomal form is released extracellularly in the presence of Ca(2+) and promotes endocytosis and plasma membrane repair (PubMed:20530211). Sphingomyelin phosphodiesterase, processed form. This form is generated following cleavage by CASP7 in the extracellular milieu in response to bacterial infection (PubMed:21157428). It shows increased ability to convert sphingomyelin to ceramide and promotes plasma membrane repair (By similarity). Plasma membrane repair by ceramide counteracts the action of gasdermin-D (GSDMD) perforin (PRF1) pores that are formed in response to bacterial infection (By similarity). (Microbial infection) Secretion is activated by bacteria such as P. aeruginos, N. gonorrhoeae and others, this activation results in the release of ceramide in the outer leaflet of the plasma membrane which facilitates the infection. (Microbial infection) Secretion is activated by human coronaviruses SARS-CoV and SARS-CoV-2 as well as Zaire ebolavirus, this activation results in the release of ceramide in the outer leaflet of the plasma membrane which facilitates the infection. Isoform 2. Lacks residues that bind the cofactor Zn(2+) and has no enzyme activity. Isoform 3. Lacks residues that bind the cofactor Zn(2+) and has no enzyme activity.

Alternative names

What's included?

200 Test
Components
AbBlue Indicator
1 x 1 Vial
Assay Buffer
1 x 20 mL
DMSO
1 x 200 µL
Enzyme Mix
2 x 1 Vial
SMase Reaction Buffer
1 x 10 mL
Sphingomyelin
1 x 100 µL
Sphingomyelinase Standard
1 x 0.2 Unit

Recommended products

Sphingomyelinase Assay Kit (Colorimetric) (ab138876) provides a sensitive method for detecting neutral SMase activity or screening its inhibitors.

Key facts

Detection method
Colorimetric
Sample types
Cell culture extracts, Whole Blood
Assay type
Enzyme activity
Reactive species
Mammals
Assay Platform
Microplate reader
Sensitivity
= 0.08 mU/mL

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Notes

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

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.

Associated diseases and disorders

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.

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  • Functional Studies - Sphingomyelinase Assay Kit (Colorimetric) (ab138876), expandable thumbnail

    Functional Studies - Sphingomyelinase Assay Kit (Colorimetric) (ab138876)

    Sample Standard Curve for Sphingomyelinase. Sphingomyelinase dose response was measured on a 96-well plate with ab138876 using a microplate reader. As low as 0.08 mU/ml sphingomyelinase can be detected with 60 minutes incubation (n=3).

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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