C12 0 sphingomyelin

Manufactured by Avanti Polar Lipids

Sourced in United States

C12:0-sphingomyelin is a synthetic lipid compound produced by Avanti Polar Lipids. It is a sphingolipid with a saturated 12-carbon fatty acid chain. The primary function of C12:0-sphingomyelin is to serve as a research tool and structural component in the study of lipid membranes and biological systems.

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Lab products found in correlation

C17 sphingosine, by Avanti Polar Lipids (3 mentions) C17 sphinganine, by Avanti Polar Lipids (3 mentions) C25 0 ceramide, by Avanti Polar Lipids (2 mentions) C12 0 ceramide, by Avanti Polar Lipids (2 mentions) Enzymatic colorimetric assay, by Fujifilm (1 mentions) Sphingomyelin, by Avanti Polar Lipids (1 mentions)

Close spelling variants of this product

Sphingomyelin (65 citations)

4 protocols using c12 0 sphingomyelin

Extraction and Quantification of Sphingolipids

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To measure sphingolipids, total lipids were extracted as previously described (19 (link)). Briefly, cell pellets were re-suspended in chloroform/methanol/water (10:5:1; v/v/v), followed by addition of internal standards containing 0.5 nmoles of C12:0 ceramide, C17-sphingosine, C17-sphinganine, C25:0-ceramide and C12:0-sphingomyelin from Avanti Polar Lipids (Alabaster, AL). After tip sonication, samples were incubated at 48 °C overnight. The next day, 100 μl of aliquots were taken out and dried under N₂ for measurement of total phospholipids (Wako Chemicals, Neuss, Germany). The rest samples were added with 75 μl of 1M KOH in methanol, sonicated for 30 min, then incubated at 37 °C for 2 h and dried in a nitrogen evaporator.

WANG Y., PARK N.Y., JANG Y., MA A, & JIANG Q. (2015). Vitamin E γ-tocotrienol inhibits cytokine-stimulated NF-κB activation by induction of anti-inflammatory A20 via stress adaptive response due to modulation of sphingolipids. Journal of immunology (Baltimore, Md. : 1950), 195(1), 126-133.

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Sphingolipid Extraction and Quantification

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To measure sphingolipids, total lipids were extracted as previously described [19 (link)]. Briefly, cell pellets were re-suspended in methanol/chloroform /water (10:5:1 [v/v/v]), followed by addition of internal standards containing 0.5 nmoles of C12:0-ceramide (Cer), C17-sphingosine, C17sphinganine, C25:0-Cer, and C12:0-sphingomyelin from Avanti Polar Lipids (Alabaster, AL). After tip sonication, samples were incubated at 48°C overnight. The next day, 100 μl aliquots was taken out and dried under N₂ for measurement of total phospholipids (Wako Chemicals, Neuss, Germany). For the rest of the samples, 75 μl of 1 M KOH in methanol was added, sonicated for 30 min, and then incubated at 37°C for 2 h and dried in a nitrogen evaporator.

YANG C, & JIANG Q. (2018). Vitamin E δ-tocotrienol inhibits TNF-α-stimulated NF-κB activation by up-regulation of anti-inflammatory A20 via modulation of sphingolipid including elevation of intracellular dihydroceramides. The Journal of nutritional biochemistry, 64, 101-109.

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Lipid Extraction and Quantification

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Lipid was extracted as previously described [23 (link)]. Briefly, cell pellets were resuspended in 500 µL of methanol, 250 µL of chloroform and 50 µL of water after the addition of 20 µL of internal standard mixture containing 25 µM of C_12:0-ceramide, C_25:0-ceramide, C₁₇-sphingosine, C₁₇-sphinganine, and C_12:0-sphingomyelin (Avanti Polar Lipids). The suspension was dispersed fully by tip sonication for 20 sec and then incubated overnight at 48 °C. 100 µL of solution was used to determine the amount of total choline-containing phospholipids (PCs) by an enzymatic colorimetric assay (Wako chemicals, Osaka, Japan) [21 (link)]. 75 µL of 1 M potassium hydroxide in methanol was added to the rest of the solution and sonicated for 30 min. Samples were then incubated at 37 °C for 2 h and evaporated under N₂.

Jang Y., Park N.Y., Rostgaard-Hansen A.L., Huang J, & Jiang Q. (2016). Vitamin E metabolite 13’-carboxychromanols inhibit pro-inflammatory enzymes, induce apoptosis and autophagy in human cancer cells by modulating sphingolipids and suppress colon tumor development in mice. Free radical biology & medicine, 95, 190-199.

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Quantification of Brain Lipids by LC-MS/MS

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Lipids were quantified using Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS). Brain tissue samples were accurately weighed, i.e. 20 mg wet weight, and lipids were extracted from brain tissues according to a previously published one-phase extraction protocol [25 (link)]. Lipids were resolved and quantified by LC-MS/MS, as described previously [26 (link)], with the exception that 1250 rather than 250 pmoles C12:0 sphingomyelin (Avanti Polar Lipids, AL, USA) internal standard was added to each sample. C12:0 sulfatide and C12:0 galactosylceramide internal standards were added at 250 pmoles/sample. Data processing was carried out using the MMSAT program [26 (link)] and the detected sphingomyelin and sulfatide species were verified as conforming to a previously identified quadratic elution profile [27 (link)]. Lipids, expressed as ratios to the relevant internal standard, were quantified using standard curves prepared with sphingomyelin, galactosylceramide, and sulfatide external standards (Avanti Polar Lipids).

Don A.S., Hsiao J.H., Bleasel J.M., Couttas T.A., Halliday G.M, & Kim W.S. (2014). Altered lipid levels provide evidence for myelin dysfunction in multiple system atrophy. Acta Neuropathologica Communications, 2, 150.

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