At 05:33 PM 10/9/96 GMT, Florence Verrier wrote:
>>I am trying to isolate eucaryotic cell membranes (e.g. BHK cells) before
>extraction of membrane proteins. For that reason I would like to get a
>convinient protocoll to isolate the whole membranes without other
>intra-cellular proteins. I tried to find a good protocoll, but so far I
>was not satisfied. So I wonder if there might be a colluege out there who
>has done anything similar and would share his/her protocoll with me, or
>would give me some references.
>Thank you very much in advance for your help. Could you please send any
>suggestions directly to my e-mailadress:
>fverrier at pasteur.fr>>Best regards, FV
I have routinely prepared membrane fractions from plant cells using Sucrose
density gradients. I centrifuge 18,000xg supernatant at 120,000xg and pellet
all the membranes. This then may be loaded on to a sucrose step gradient
(works better than than isocratic (or continuous) gradients) for further
fractionation. Get hold of the Meth. Enzymol series from your library and
look the protocol up.
Dr. Hiranya Sankar Roychowdhury
Plant Genetic Engineering Lab.
New Mexico State University
Las Cruces, NM 88003
Ph. (505) 646-5785
hroychow at nmsu.edu