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resuspend membrane pellet


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#1 genz

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Posted 19 March 2009 - 10:25 PM

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

#2 molgen

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Posted 22 March 2009 - 01:04 AM

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.

#3 jiq724

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Posted 06 May 2009 - 07:12 AM

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

#4 klinmed

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Posted 06 May 2009 - 08:51 AM

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) ?




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