Comprehensive Sphingolipid Analysis Protocol

FcR Blocking Reagent, Viobility™ 488/520 Fixable Dye, Ly-6G VioBlue^® (REAfinity™, clone REA526), CD45 VioGreen^® (REAfinity™, clone REA737), CD11c PE-Vio^® 770 (clone REA754), and F4/80 APC (clone REA126) were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany). Zymosan was obtained from InvivoGen (Toulouse, France). All other analytes and reagents were purchased from Sharlab (Sharlab S.L., Sentmenat, Spain), or Sigma-Aldrich (Sigma-Aldrich Chemical Co., Saint Quentin Fallavier, France). All the reagents were HPLC analytical grade, except those used for UHPLC-MSMS, which were of LC-MS grade. The 10 sphingolipids used as internal standards obtained from Sigma-Aldrich were manufactured by Avanti Polar Lipids under the trade name “Ceramide/Sphingoid Internal Standard Mixture I” and corresponded to C17 sphingosine (d17:1), C17 sphinganine (d17:0), C17 sphingosine-1-P (d17:1P), C17 sphinganine-1-P (d17:0P), lactosyl (ß) C12 ceramide (Lac18:1/12:0), C12 sphingomyelin (SM18:1/12:0), glucosyl (ß) C12 ceramide (Glu18:1/12:0), 12:0 ceramide (18:1/12:0), 12:0 ceramide-1-P (18:1/12:0P), and 25:0 ceramide (18:1/25:0) in ethanol solution. The 33 sphingolipids used as standards obtained from Sigma-Aldrich were solubilized in ethanol at a concentration of 25 µM and corresponded to deoxysphingosine (dSo = m18:1); deoxysphinganine (dSa = m18:0); sphingosine (So = d18:1); sphinganine (Sa = d18:0); sphingosine-1-P (d18:1P); sphinganine-1-P (d18:0P); glucosylsphingosine (GluSo); lysosphingomyelin (LysoSM); lactosylsphingosine (LacSo); N-acetylsphingosine (18:1/2:0); N-acetylsphinganine (18:0/2:0); ceramides: 18:1/14:0, 18:1/16:0, 18:1/18:0, 18:1/20:0, 18:1/22:0, 18:1/24:1, and 18:1/24:0; ceramide-1P: 18:1/16:0P; dihydroceramides: 18:0/16:0, and 18:0/24:0; glucosylceramides: Glu18:1/16:0 and Glu18:1/24:1; lactosylceramides: Lac18:1/16:0 and Lac18:1/24:1; sphingomyelins: SM18:1/14:0, SM18:1/16:0, SM18:1/18:0, SM18:1/18:1, SM18:1/20:0, SM18:1/22:0, SM18:1/24:1, and SM18:1/24:0.

Free full text: Click here

Pierron A., Guzylack-Piriou L., Tardieu D., Foucras G, & Guerre P. (2023). Zymosan-Induced Murine Peritonitis Is Associated with an Increased Sphingolipid Synthesis without Changing the Long to Very Long Chain Ceramide Ratio. International Journal of Molecular Sciences, 24(3), 2773.

Publication 2023

Ceramide Clone Dihydroceramides Ethanol Glucosylceramides Glucosylsphingosine Hplc Lactosylceramides Lactosylsphingosine Lipids Lysosphingomyelin N acetylsphingosine Reagent Sphinganine Sphingolipids Sphingomyelins Sphingosine Zymosan

Corresponding Organization : Université de Toulouse

Top 5 similar protocols

Variable analysis

independent variables

FcR Blocking Reagent
Zymosan

dependent variables

Ly-6G VioBlue® (REAfinity™, clone REA526)
CD45 VioGreen® (REAfinity™, clone REA737)
CD11c PE-Vio® 770 (clone REA754)
F4/80 APC (clone REA126)

control variables

Viobility™ 488/520 Fixable Dye
10 sphingolipids used as internal standards
33 sphingolipids used as standards

positive controls

Zymosan

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!