Lipids were extracted from mouse brain (500 mg) with 9 ml chloroform–methanol (2:1, v/v) and then with 7.6 ml chloroform–methanol-water (1:2:0.8, v/v/v).
The extraction was dried under a gentle stream of nitrogen and re-dissolved with 7.5 ml chloroform-methanol-water(30:60:8; v/v/v).
Samples were applied to a DEAE-Sephadex A-25 column (6 x 45 mm, acetate form) (Amersham Biosciences AB, Sweden).
Neutral lipids were eluted with 15 ml chloroform-methanol-water (30:60:8; v/v/v).
Acidic glycolipid mixtures were eluted with 7.5 ml chloroform-methanol-0.8 M sodium acetate (30:60:8; v/v/v).