Removing the lipid from the protein sample

Frank O. Fackelmayer fof1 at chclu.chemie.uni-konstanz.de
Fri Dec 13 11:49:43 EST 1996


dykim at bioneer.kaist.ac.kr (Dooyeon Kim) wrote:
>Hi, Netters. Please help me to remove the too much lipid in my samples. 
>I was trying to show the clear protein bands analyzed by SDS-PAGE with
>my protein samples. The problem is because my sample contains the too
>much lipids (it is the proteoliposomes),most of the bands are streaked
>or pushed upwards. To remove the lipids, I tried to use the TCA-acetone
>precipitation but could not get good results.

The method Dr E.Buxbaum talkes about is good and reliable. However, it is
not a real precipitation as your protein remains in a small volume of
water. Of course, you can dry it down but that takes some time. This is
why I prefer to use the method by Wessel & Fluegge (1984), Anal. Biochem.
138:141-143. It´s a methanol/chloroform precipitation and gives you a
pellet that is easily redissolved. The method was especially devised for
removing lipids or detergents, so it should be perfect for you. 

Hope it helps,
Frank

-- 
Dr. Frank O. Fackelmayer
Division of Biology
University of Konstanz
D-78434 Konstanz
Germany



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