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,
Dr. Frank O. Fackelmayer
Division of Biology
University of Konstanz
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