Phospholipids and their hydroxy and hydroperoxy derivatives, including 15-HETE-PE and 15-HpETE-PE, were analyzed by LC/MS using a Dionex Ultimate 3000 HPLC system coupled on-line to an Orbitrap Fusion Lumos mass spectrometer (Thermo Fisher Scientific) using a normal phase column (Luna 3 m silica C18(2) 100 , 150 2.0 mm, Phenomenex)
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Boyd A, Thomas R, Hansell AL, Gulliver J, Hicks LM, Griggs R, et al
Then pellets were resuspended in lysis buffer (Tris-HCl 100 mM and 0.5% Triton X-100, pH 8) containing Halt Protease and Phosphatase Inhibitor cocktail (Thermo Scientific, 1861280), thoroughly vortexed for 1 min and incubated for 15 min on ice