Fig. 3. TLC analysis of the lipid composition of the amidohydrolase assay with FAAH protein and different alkamide substrates (1–12). Reactions were initiated by the addition of purified FAAH protein (0.9 µg) with 50 mM Bis-Tris propane–HCl, (pH 9.0), at 30 °C for 30 min, 120 rpm, and 100 µM of different alkamides or NAE 12:0 in a final volume of 0.45 ml. Lipids were extracted and then separated by TLC (hexane/Et2O/AcOH, 80:20:2, v/v/v). Lipids were visualized by spraying the plate with a solution of primuline and exposure under a UV lamp (Testet et al., 2005). Position of the origin, free lauric acid formed by hydrolysis of NAE 12:0 and free fatty acid (FFA) 12:0 standard are indicated (arrows).