1 14c fatty acids

Acyl-CoAs ([1-¹⁴ C]-labeled an unlabeled) used for enzymatic assays were synthesized according to the method developed by Sánchez et al. [47 (link)]. [1-¹⁴C]-fatty acids and triolein [carboxyl-¹⁴C] were purchased from Perkin Elmer or American Radiochemicals. Labeled diacylglycerol was derived through partial lipase treatment of triolein [carboxyl-¹⁴C] with Rhizopus arrhizus lipase (Sigma-Aldrich). The lipase products were separated on TLC plates (silica gel 60; Sigma-Aldrich). The DAG was eluted from the silica with methanol: chloroform (2:1, v/v), extracted with chloroform by Bligh and Dyer method (1959). Non-radioactive fatty acids and diacylglycerol were ordered from Larodan or Sigma-Aldrich. Internal standard of heptadecanoic acid methyl ester, coenzyme A and TAG standard for thin-layer chromatography were purchased also from Sigma-Aldrich.

Fatty acids and Coenzyme A were obtained from Nu-Chek Prep Inc (Elysian, MN) and from Larodan (Malmö, Sweden), [1-¹⁴C]Fatty acids from PerkinElmer or Lipidox (Stockholm, Sweden). Palmitoyl(16:0)-lysophosphatidylcholine (16:0-lysoPtdCho), essentially fatty acid free bovine serum albumin (BSA), and other fine chemicals were purchased from Sigma-Aldrich (St. Louis, USA). [¹⁴C]Acyl-CoA and acyl-CoAs were synthesized according to the method described by [19 (link)]. Argentation TLC plates were prepared by immersing TLC plates (silica 60, Merck) in AgNO₃ (12%, w/v) dissolved acetone/water (4:1 by vol.) and drying the plates at room temperature.