Overview

  • Product name

    Sphingomyelinase Assay Kit (Colorimetric)
    See all Sphingomyelinase kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture extracts, Whole Blood
  • Assay type

    Enzyme activity
  • Sensitivity

    0.08 mU/ml
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    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.

  • Notes

    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.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 200 tests
    AbBlue Indicator 1 vial
    Assay Buffer 1 x 20ml
    DMSO 1 x 200µl
    Enzyme Mix 2 vials
    SMase Reaction Buffer 1 x 10ml
    Sphingomyelin 1 x 100µl
    Sphingomyelinase Standard 1 x 0.2 unit
  • Research areas

  • Function

    Converts sphingomyelin to ceramide. Also has phospholipase C activities toward 1,2-diacylglycerolphosphocholine and 1,2-diacylglycerolphosphoglycerol. Isoform 2 and isoform 3 have lost catalytic activity.
  • Involvement in disease

    Defects in SMPD1 are the cause of Niemann-Pick disease type A (NPDA) [MIM:257200]; also known as Niemann-Pick disease classical infantile form. It is an early-onset lysosomal storage disorder caused by failure to hydrolyze sphingomyelin to ceramide. It results in the accumulation of sphingomyelin and other metabolically related lipids in reticuloendothelial and other cell types throughout the body, leading to cell death. Niemann-Pick disease type A is a primarily neurodegenerative disorder characterized by onset within the first year of life, mental retardation, digestive disorders, failure to thrive, major hepatosplenomegaly, and severe neurologic symptoms. The severe neurological disorders and pulmonary infections lead to an early death, often around the age of four. Clinical features are variable. A phenotypic continuum exists between type A (basic neurovisceral) and type B (purely visceral) forms of Niemann-Pick disease, and the intermediate types encompass a cluster of variants combining clinical features of both types A and B.
    Defects in SMPD1 are the cause of Niemann-Pick disease type B (NPDB) [MIM:607616]; also known as Niemann-Pick disease visceral form. It is a late-onset lysosomal storage disorder caused by failure to hydrolyze sphingomyelin to ceramide. It results in the accumulation of sphingomyelin and other metabolically related lipids in reticuloendothelial and other cell types throughout the body, leading to cell death. Clinical signs involve only visceral organs. The most constant sign is hepatosplenomegaly which can be associated with pulmonary symptoms. Patients remain free of neurologic manifestations. However, a phenotypic continuum exists between type A (basic neurovisceral) and type B (purely visceral) forms of Niemann-Pick disease, and the intermediate types encompass a cluster of variants combining clinical features of both types A and B. In Niemann-Pick disease type B, onset of the first symptoms occurs in early childhood and patients can survive into adulthood.
  • Sequence similarities

    Belongs to the acid sphingomyelinase family.
    Contains 1 saposin B-type domain.
  • Cellular localization

    Lysosome.
  • Information by UniProt
  • Alternative names

    • Acid sphingomyelinase
    • ASM
    • ASM_HUMAN
    • aSMase
    • NPD
    • Smpd1
    • Sphingomyelin phosphodiesterase
    • Sphingomyelin phosphodiesterase 1 acid lysosomal
    see all

Images

  • 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).

Protocols

References

This product has been referenced in:

See all 2 Publications for this product

Customer reviews and Q&As

Question
Answer

The sphingomyelin substrate in the sphingomyelinase assay ab138876 is from chicken egg and is assumed to be a mixture, not a single sphingomyelin of known length.

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