method for membrane receptor extraction

WS via methods%40net.bio.net (by novalidaddress from nurfuerspam.de)
Wed Mar 3 09:15:20 EST 2010


Hi Ketut,

if you can't find any special procedure for your receptor in the
literature, I'd try simple fractionation:

first, disrupt your cells in a buffer that does not contain
detergents. This will remove the cytosol. Remainder should be able for
recovery at 15.000g. Wash the pellet a few times with this first
buffer. Then extract the pellet with a buffer containing 0.5 to 1% of
nonionic detergent. This should solubilize most of the membrane
proteins, except those from resistant structures like the lipid rafts.
When you spin again, you will sediment the nuclei.

HTH,

Wo


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