Overview

  • Product name

    Sphingomyelin Assay Kit (Fluorometric)
    See all Sphingomyelin kits
  • Detection method

    Fluorescent
  • Sample type

    Cell culture extracts, Whole Blood
  • Assay type

    Quantitative
  • Sensitivity

    1 µM
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Abcam's Sphingomyelin Assay Kit (Fluorometric) (ab138877) provides the most sensitive method for detecting neutral SM activity or screening SM inhibitors. The kit uses AbRed Indicator as a fluorogenic probe to indirectly quantify the phosphocholine produced from the hydrolysis of sphingomyelin (SM) by sphingomyelinase (SMase). It can be used for measuring the amount of SM in blood, cell extracts or other solutions. The fluorescence intensity of AbRed Indicator is proportional to the formation of phosphocholine, therefore to the amount of SM. AbRed Indicator enables the assay readable by either a fluorescence reader or an absorbance reader.

    The kit is an optimized “mix and read” assay that can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step.

  • Notes

    Sphingomyelin (SM) is largely found in the exoplasmic leaflet of the cell membrane, primarily in nervous tissue. It plays an important role in signal transduction. Sphingomyelin accumulates abnormally in Niemann-Pick disease and Abetalipoproteinemia.

  • Platform

    Microplate reader

Properties

Images

  • Sphingomyelin dose response was measured on a solid black 96-well plate with ab138877 using a fluorescence microplate reader. As low as 1 µM sphingomyelin can be detected with 60 minutes incubation (n=3).

Protocols

References

This product has been referenced in:

  • Im EJ  et al. Sulfisoxazole inhibits the secretion of small extracellular vesicles by targeting the endothelin receptor A. Nat Commun 10:1387 (2019). Read more (PubMed: 30918259) »
  • Aligeti M  et al. Cooperation between the Hepatitis C Virus p7 and NS5B Proteins Enhances Virion Infectivity. J Virol 89:11523-33 (2015). Read more (PubMed: 26355084) »
See all 2 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

1. How do I prepare cell lysates for this assay?
We would recommend you to use our Mammalian Cell Lysis Buffer 5X (ab179835) to lyse the cells and use the supernatant.


2. Is this assay sensitive to detergents and solvents?
Yes, it is sensitive to Tween 20 which will increase background dramatically even at 0.1%, both 0.1% Triton X 100 and SDS are OK.


3. What is the linear range for the assay?
0.3-30 uM (best fit 1-10 uM)


Read More

Answer

Thank you for contacting us.


We don't have experience using this kit on tissue sample, however the following general protocol should be suitable.



1. Homogenize ˜200mg/ml of the brain tissue in PBS.

2. Centrifuge at 800g for 10 min at 4C.

3. Transfer the supernatant to another tube and sonicate the supernatant 20X at one second bursts.

4. Use the supernatant for the assay, freeze unused sample at-80C for later use, the sample should be stable for 2-3 weeks.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

-----------------------------------------------------------------------------------------------

Can’t find the antibody you need? Try our Custom RabMAb® services.

Discover More at www.abcam.com/tag/custom%20services

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up