Amplite® Fluorimetric Sphingomyelin Assay Kit *Red Fluorescence*

Amplite® Fluorimetric Sphingomyelin Assay Kit

Red Fluorescence

Amplite® Fluorimetric Sphingomyelin Assay Kit *Red Fluorescence* is a fluorimetric assay kit for quantification of sphingomyelin in blood, cell extracts, and other solutions.

Detection Principle: Quantifies phosphocholine generated from sphingomyelin hydrolysis by sphingomyelinase.
Detection Chemistry: Uses Amplite® Red as a fluorogenic probe for sphingomyelin measurement.
Signal Response: Fluorescence intensity is proportional to the amount of sphingomyelin in the sample.
Workflow: Optimized mix-and-read assay compatible with 96- or 384-well microplates and automation without a separation step.
Instrument Compatibility: Fluorescence detection at Ex/Em 540/590 nm using solid black microplates.

Sphingomyelin dose response was measured on a solid black 96-well plate with Amplite® Fluorimetric Sphingomyelin Assay Kit using a Gemini fluorescence microplate reader (Molecular Devices).

Protocol

SDS

Catalog	Size	Price	Quantity
13625	100 Tests	Price

Overview

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. It accumulates abnormally in Niemann-Pick disease and Abetalipoproteinemia. Our Amplite® Fluorimetric Sphingomyelin Assay Kit provides the most sensitive method for detecting neutral SM activity or screening its inhibitors. The kit uses Amplite® Red 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 SM in blood, cell extracts or other solutions. The fluorescence intensity of Amplite® Red is proportional to the formation of phosphocholine, therefore to the amount of SM. Amplite® Red enables the assay readable either in fluorescence intensity or absorption mode. 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.

Citations

View all 10 citations: Citation Explorer

St-N, a novel alkaline derivative of stevioside, reverses docetaxel resistance by targeting lysosomes in vitro and in vivo

Authors:

Guo, Yanxia and Wang, Shikang and Liu, Qun and Dong, Yan and Liu, Yongqing

Journal:

PloS one (2024): e0316268

Sphingomyelin synthase 1 supports two steps of rubella virus life cycle

Authors:

Yagi, Mayuko and Hama, Minami and Ichii, Sayaka and Nakashima, Yurie and Kanbayashi, Daiki and Kurata, Takako and Yusa, Kosuke and Komano, Jun

Journal:

iScience (2023)

Polyphyllin D “Punctures” Hypertrophic Lysosomes to Reverse Drug Resistance of Hepatocellular Carcinoma by Targeting Acid Sphingomyelinase

Authors:

Wang, Yang and Chen, Yan-Yan and Gao, Gui-Bin and Zheng, Yang-Han and Yu, Nan-Nan and Ouyang, Lan and Gao, Xuejuan and Li, Nan and Wen, Shi-Yuan and Huang, Shangjia and others,

Journal:

Molecular Therapy (2023)

Sphingomyelin synthase activity affects TRIF-dependent signaling of Toll-like receptor 4 in cells stimulated with lipopolysaccharide

Authors:

Prymas, Kamila and Swiatkowska, Anna and Traczyk, Gabriela and Ziemli&nacute;ska, Ewelina and Dziewulska, Anna and Ciesielska, Anna and Kwiatkowska, Katarzyna

Journal:

Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids (2019): 158549

Doxepin mitigates noise induced neuronal damage in primary auditory cortex of mice via suppression of acid sphingomyelinase/ceramide pathway

Authors:

Su, Yu-Ting and Meng, Xing-Xing and Zhang, Xi and Guo, Yi-Bin and Zhang, Hai-Jun and Cheng, Yao-Ping and Xie, Xiao-Ping and Chang, Yao-Ming and Bao, Jun-Xiang

Journal:

The Anatomical Record (2017)

References

View all 65 references: Citation Explorer

Expression of alkaline sphingomyelinase in yeast cells and anti-inflammatory effects of the expressed enzyme in a rat colitis model

Authors:

Andersson D, Kotarsky K, Wu J, Agace W, Duan RD.

Journal:

Dig Dis Sci (2009): 1440

A novel sphingomyelinase-like enzyme in Ixodes scapularis tick saliva drives host CD4 T cells to express IL-4

Authors:

Alarcon-Chaidez FJ, Boppana VD, Hagymasi AT, Adler AJ, Wikel SK.

Journal:

Parasite Immunol (2009): 210

A novel mitochondrial sphingomyelinase in zebrafish cells

Authors:

Yabu T, Shimuzu A, Yamashita M.

Journal:

J Biol Chem (2009): 20349

Alterations of myelin-specific proteins and sphingolipids characterize the brains of acid sphingomyelinase-deficient mice, an animal model of Niemann-Pick disease type A

Authors:

Buccinna B, Piccinini M, Prinetti A, Sc and roglio F, Prioni S, Valsecchi M, Votta B, Grifoni S, Lupino E, Ramondetti C, Schuchman EH, Giordana MT, Sonnino S, Rinaudo MT.

Journal:

J Neurochem (2009): 105

A complex extracellular sphingomyelinase of Pseudomonas aeruginosa inhibits angiogenesis by selective cytotoxicity to endothelial cells

Authors:

Vasil ML, Stonehouse MJ, Vasil AI, Wadsworth SJ, Goldfine H, Bolcome RE, 3rd, Chan J.

Journal:

PLoS Pathog (2009): e1000420

Spectral properties

Excitation (nm)	571
Emission (nm)	584

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Fluorescence microplate reader
Excitation	540 nm
Emission	590 nm
Cutoff	570 nm
Recommended plate	Solid black

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Fax
Email	sales@aatbio.com
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Bulk request	Inquire
Custom size	Inquire
Technical support	Contact us
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Purchase order	Send to sales@aatbio.com
Shipping	Standard overnight for United States, inquire for international

Page updated on October 8, 2026

Component A: Enzyme Mix	1 bottle (lyophilized powder)
Component B: Sphingomyelinase	1 vial (lyophilized powder)
Component C: Amplite® Red	1 vial (lyophilized powder)
Component D: SMase Reaction Buffer	1 bottle (20 mL)
Component E: Assay Buffer	1 bottle (5 mL)
Component F: 50 mM Sphingomyelin Standard	1 vial (20 µL )
Component G: DMSO	1 vial (200 µL)