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resuspend membrane pellet - (Mar/19/2009 )

Hi everyone,

I am working with a membrane protein. Fractionation involves a centrifugation step at 100 000 g for 1 hour. However, the pellet after centrifugation is so resilient to resuspension.

Can anyone suggest a good way to resuspend a membrane protein pellet after high speed spin?
Thanks inadvance

GE

-genz-

Do you need it native? Can you denature it?

Membrane proteins have hydrophobic properties. A detergent can help you suspend them but choose it well according to your downstream applications.

-molgen-

my protein could be sonicated in sucrose buffer. Maybe that would be help.

-jiq724-

genz on Mar 20 2009, 08:25 AM said:

Hi everyone,

I am working with a membrane protein. Fractionation involves a centrifugation step at 100 000 g for 1 hour. However, the pellet after centrifugation is so resilient to resuspension.

Can anyone suggest a good way to resuspend a membrane protein pellet after high speed spin?
Thanks inadvance

GE

I assume from speed/time that you are using centrifugation to concentrate protein-containing membranes and not free protein.

Why not avoid the pellet resuspension problem entirely and spin the membranes onto a sucrose cushion (say 1.5 . 2 M) ?

-klinmed-