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Active Acid Sphingomyelinase (ASM)

SMPD1; NPD; SMase; aSMase; Sphingomyelin Phosphodiesterase 1,Acid Lysosomal; Simply Sphingomyelinase

ACTIVITY TEST

Sphingomyelin phosphodiesterase 1 (SMPD1), also known as acid sphingomyelinase (ASM), belongs to the sphingomyelin phosphodiesterase family. The protein Converts sphingomyelin to ceramide. ASM also has phospholipase C activities toward 1,2-diacylglycerolphosphocholine and 1,2-diacylglycerolphosphoglycerol.
Thus, the recombinant human ASM activity was measured by its ability to hydrolyze 2-N-Hexadecanoylamino-4-nitrophenylphosphorylcholine (HDA-PC) to p-Nitrophenol. The reaction was performed in 50 mM MES, 0.5 µM ZnCl2, pH 7.5 ( Assay Buffer), initiated by addition 50 μL of various concentrations of ASM (diluted with Assay Buffer) to 50 µL of 1 mM Substrate HDA-PC ( 50 mM stock solution in methanol, diluted with Assay Buffer). Incubated at room temperature for 20 minutes in the dark, then read at a wavelength of 405 nm.

One unit of enzyme activity is defined as the 1μg of enzyme required to convert 1pmol of HDA-PC to p-Nitrophenol in 1min at 37°C. The specific activity of recombinant human ASM is >1000 pmol/min/µg.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Blood.? Acid sphingomyelinase is activated in sickle cell erythrocytes and contributes to inflammatory microparticle generation in SCD Pubmed:25075126
Journal of Clinical Lipidology Increase in acid sphingomyelinase level in human retinal endothelial cells and CD34+ circulating angiogenic cells isolated from diabetic individuals is associated with dysfunctional retinal vasculature and vascular repair process in diabetes 10.1016/j.jacl.2017.03.007
Diabetes & metabolism Fenofibrate decreases plasma ceramide in type 2 diabetes patients: A novel marker of CVD? pubmed:28499696
INFLAMMATION Acid Sphingomyelinase and Acid ¦Â-Glucosidase 1 Exert Opposite Effects on Interleukin-1¦Â-Induced Interleukin 6 Production in Rheumatoid Arthritis Fibroblast-Like Synoviocytes 33665756
Indian J Ophthalmol Evaluation of sphingolipid metabolism on diabetic retinopathy 34708809
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