On Mon, 11 Aug 2003, Greg Pankhurst wrote:
> The problem is getting rid of all the lipid from the membranes.
I used to delipidate samples for IEF, as follows:
- Get the protein as a pellet
- Add 500 ul organic solvent
- Resuspend by sonicating on ice
- Precipitate for 30 min on ice
- Pellet by centrifugation at 13000 rpm x 5 min
Nice and easy!
I used acetone as a solvent, but i have since heard that ether is better,
as very hydrophobic proteins are less soluble in it. I was working on the
whole proteome at the time, so whilst i did get some membrane proteins
out, i can't say whether this is a good technique for membrane proteins in
particular.
tom
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REMOVE AND DESTROY
