Delipidation of Membrane Fraction for HPLC Analysis

Tom Anderson univ0938 at herald.ox.ac.uk
Mon Aug 11 07:20:29 EST 2003

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



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