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3 protocols using dilauroyl phosphatidylcholine

1

Lipid Monolayer Characterization in Saline

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Dilauroyl phosphatidylcholine (DLPC), dipalmitoyl phosphatidylcholine (DPPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), the lipophilic uorescent probe L,a- phosphatidylethanolamine-N-lissamine rhodamine B sulfonylammonium salt and chicken egg-transphosphatidylated (RhhoPEegg) were purchased from Avanti Polar Lipids. DiIC18 (1,1 0 -dioctadecyl-3,3,3 0 ,3 0 -tetramethylindocarbocyanine perchlorate) was purchased from Molecular Probes and dihydrocholesterol (dchol, a less hydrolyzable cholesterol analog 22 ) was obtained from Sigma. 1-Palmitoyl-2-lauroyl-sn-glycero-3phosphocholine was a generous gi from Dr Bruno Maggio.
Solvents and chemicals were of the highest commercial purity available. The water used for the subphase was from a Milli-Q system (Millipore), 18 MU cm and lipid monolayers were prepared and characterized on subphases of 0.15 M NaCl at 21 AE 1 C.
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2

Lipid Extraction for Mass Spectrometry

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For lipid compound analysis, 5 mg of tissue was homogenized with 225 μL of MeOH and 25 μL of 500 μg/L dilauroylphosphatidylcholine (PC 12:0/12:0; Avanti Polar Lipids, AL, USA) dissolved in MeOH as the internal standard. After being left on ice, the solution was centrifuged at 16,000×g for 5 min at 4 °C, and The extracted solutions were subjected to LC/MS analysis.
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3

Lipid Sourcing and Preparation

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Dilauroyl phosphatidylcholine (DLPC), dilauroyl phosphatidylglycerol (DLPG), palmitoyl oleoyl phosphatidylcholine (POPC), and palmitoyl oleoyl phosphatidylglycerol (POPG) were purchased from Avanti® Polar Lipids, Inc. Monosodium phosphate and disodium phosphate were purchased from Sigma-Aldrich.
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