Name: Amplite® Fluorimetric Acidic Sphingomyelinase Assay Kit *Red Fluorescence*
Brand: AAT Bioquest
SKU: 13622
Price: 355 USD
Availability: InStock

Amplite® Fluorimetric Acidic Sphingomyelinase Assay Kit

Red Fluorescence

Amplite® Fluorimetric Acidic Sphingomyelinase Assay Kit *Red Fluorescence* is a fluorimetric assay kit for quantification of acidic sphingomyelinase activity in blood, cell extracts, and other solutions.

Detection Principle: Indirectly quantifies phosphocholine from sphingomyelin hydrolysis.
Chemistry: Uses Amplite™ Red as the fluorogenic probe.
Workflow: Optimized mix-and-read format compatible with HTS liquid handling.
Instrument Compatibility: Fluorescence detection at Ex/Em 540/590 nm.
Applications: Supports acidic SMase activity measurement and inhibitor screening.

Sphingomyelinase (from human placenta) dose response was measured on a 96-well half-area black plate with Amplite® Fluorimetric Acidic Sphingomyelinase Assay Kit (13622) using a fluorescence microplate reader. 20 µL of SMase standard or control was incubated with 20 µL of sphingomyelin working solution at  37 °C for 3 hours, and then 20 µL of sphingomyelinase assay mixture was added into each well. The signals shown in the figure are the readings at Ex/Em = 540/590 nm (cut off at 570 nm) after 2 hours incubation at room temperature.

Protocol

SDS

Catalog	Size	Price	Quantity
13622	200 Tests	Price

Overview

Sphingomyelinase (SMase) is an enzyme that is responsible for cleaving sphingomyelin (SM) to phosphocholine and ceramide. Activation of SMases plays an important role in the cellular response such as regulation of cell growth, cell differentiation, cell cycle arrest and programmed cell death. Five types of sphingomyelinase (SMase) have been identified based on their cation dependence and pH optima of action, including lysosomal acid SMase, secreted zinc-dependent acid SMase, magnesium-dependent neutral SMase, magnesium-independent neutral SMase and alkaline SMase. Among them, the lysosomal acidic SMase and the magnesium-dependent neutral SMase are considered to be the major factors for the production of ceramide in cellular stress responses. Our Amplite® Fluorimetric Acidic Sphingomyelinase Assay Kit provides one of the most sensitive methods for detecting acidic SMase 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 SMase activity in blood, cell extracts or other solutions. The fluorescence intensity of Amplite® Red is proportional to the formation of phosphocholine, therefore to the SMase activity. The kit is an optimized "mix and read" assay compatible with HTS liquid handling instruments.

Citations

View all 10 citations: Citation Explorer

Rapamycin induced autophagy enhances lipid breakdown and ameliorates lipotoxicity in Atlantic salmon cells

Authors:

Phadwal, Kanchan and Haggarty, Jennifer and Kurian, Dominic and Marti, Judit Aguilar and Sun, Jianxuan and Houston, Ross D and Betancor, M{\'o}nica B and MacRae, Vicky E and Whitfield, Phillip D and Macqueen, Daniel J

Journal:

Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids (2025): 159636

Extracellular high molecular weight $\alpha$-synuclein oligomers induce cell death by disrupting the plasma membrane

Authors:

Ito, Naohito and Tsuji, Mayumi and Adachi, Naoki and Nakamura, Shiro and Sarkar, Avijite Kumer and Ikenaka, Kensuke and Aguirre, C{\'e}sar and Kimura, Atsushi Michael and Kiuchi, Yuji and Mochizuki, Hideki and others,

Journal:

npj Parkinson's Disease (2023): 139

Sortilin deletion in the prefrontal cortex and hippocampus ameliorates depressive-like behaviors in mice via regulating ASM/ceramide signaling

Authors:

Chen, Shu-jian and Gao, Cong-cong and Lv, Qun-yu and Zhao, Meng-qi and Qin, Xiao-ying and Liao, Hong

Journal:

Acta Pharmacologica Sinica (2021): 1--15

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): 2220--2232

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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Page updated on October 8, 2026

Component A: Enzyme Mix	2 bottles (lyophilized powder)
Component B: Sphingomyelin	1 vial (100 µL)
Component C: Amplite® Red	1 vial (lyophilized powder)
Component D: SMase Reaction Buffer	1 bottle (10 mL)
Component E: Assay Buffer	1 bottle (10 mL)
Component F: DMSO	1 vial (200 µL)