Sphingomyelin Assay Kit (Fluorometric) (ab138877)

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製品の概要

  • 製品名

    Sphingomyelin Assay Kit (Fluorometric)
    Sphingomyelin キット 製品一覧

  • 検出方法

    Fluorescent

  • サンプルの種類

    Cell culture extracts, Whole Blood

  • アッセイタイプ

    Quantitative

  • 検出感度

    1 µM

  • 種交差性

    交差種: Mammals, Other species

  • 製品の概要

    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.

  • 特記事項

    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.

  • 試験プラットフォーム

    Microplate reader

製品の特性

画像

  • Sample Standard Curve for Sphingomyelin
    Sample Standard Curve for Sphingomyelin
    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).

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参考文献

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

ab138877 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

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Question

I am interested in purchasing this kit, But I have some questions. Hopefully you will be able to answer me.
1. How do I prepare cell lysates for this assay?
2. Is this assay sensitive to detergents and solvents?
3. What is the linear range for the assay?

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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)


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Question



Inquiry: We want to use ab138877 (Sphingomyelin Assay kit) kit to determinate sphingomyelin in brain (hippocampus) and we want to know ¿which could be the better way to disaggregate the tissue? ¿In wich buffer, solvent or solution? ¿Do we need to make an organic extraction? Thank you. Best regards

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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.

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