This is a cached page for the URL ( To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
Desalting Acidic Glycosphingolipids

Desalting Acidic Glycosphingolipids

Contributor: Suprya Jayadev
Date: November 10, 1993

1) Prepare a C-18 reverse phase prep-sep cartridge by washing with:

a) 10 ml of methanol

b) 20 ml of chloroform-methanol (2:1)

c) 10 ml of methanol

d) 10 ml of methanol-1.6 M sodium acetate (1:1)

2) Apply sample in 2 ml of methanol-1.6 M sodium acetate (1:1).

3) Collect wash through and apply twice more to the cartridge.

4) Wash cartridge with 40 ml of water.
---> This actually washes away the salt from the glycosphingolipids.

5) Elute acid glycosphingolipids with:

a) 2 ml of methanol

b) 10 ml of chloroform-methanol (1:1)

6) Pool the two eluates and dry lipids down.

7) Resuspend in a minimal volume of chloroform-methanol (1:1) and spot on TLC.

8) Develop TLC in chloroform-methanol-0.22% calcium chloride (60:35:8).