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2 protocols using stearic acid

1

Lipid Profiling of Immune Cells

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Monoclonal anti-ceramide antibody (mouse IgM, clone MID 15B4), ROS inhibitors APDC and BHA, and triacsin C were purchased from Sigma. All caspase inhibitors were from R&D Systems and Biovision. Ceramide synthesis inhibitor fumonisin B1 and myriocin (a serine palmitoyltransferase) were from Cayman Chemical. BODIPY-C16 FA was purchased from Thermo Fisher Scientific. All flow cytometric antibodies were purchased from Biolegend. All FA sodium salts were from Nu-Chek Prep, Inc. Palmitic acid (PA, 5mM), stearic acid (SA, 5mM) oleic acid (OA, 5mM), linoleic acid (LA, 5mM) and ω-3 eicosapentaenoic acid (EPA) were prepared with 2mM endotoxin-free BSA in PBS, sonicated until dissolved and filtered through 0.2μM sterile filter.
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2

Fatty Acid Analysis of Botanical Compounds

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TWM was obtained from Qianjin Xieli Pharmaceutical Co., Ltd (Hunan, China). TP (>98% pure) was provided by Guilin Sanjin Biologics Co., Ltd (Guanxi, China). Docosahexaenoic acid (C22:6n3), docosanoic acid (C22:0), eicosapentaenoic acid (C20:5n3), eicosatrienoic acid (C20:3n6), arachidonic acid (C20:4n6), arachidic acid (C20:0), nonadecadienoic acid (C19:2n6), γ-linolenic acid (C18:3n6), α-linolenic acid (C18:3n3), linoleic acid (C18:2n6), oleic acid (C18:1n9), cis-vaccenic acid (C18:1n7), stearic acid (C18:0), palmitic acid (C16:0), palmitoleic acid (C16:1n7), myristic acid (C14:0) and lauric acid (C12:0) were purchased from Nu-Chek Prep (Elysian, MN, USA). Their purities were above 98%. Trimethylsilane diazomethane (TMSCHN2) in n-hexane (2 mol/L) was purchased from Energy Chemical (Shanghai, China). The other chemical reagents were purchased from Xilong Scientific Co., Ltd (Guangdong, China).
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