Delipidation of Membrane Fraction for HPLC Analysis

Greg Pankhurst g.pankhurst at spamblock.cfi.unsw.edu.spamblock.au
Sun Aug 10 22:45:52 EST 2003

Hey All

I'm interested in analysing membrane proteins of various cell lines via
RP-HPLC. The plan was to mechanically lyse the cells, spin down the
membranes, solublise them and then do the analysis.

The problem is getting rid of all the lipid from the membranes. My guess
is if I inject all the lipid onto the column, all I'll succeed in doing
is binding a heap of muck to my column. I could do a TCA precipitation
of my proteins after the solubisation, problem being I'm trying to
preserve the oxidation/post translational modification state of my
proteins, and TCA precips seem cause any number of nasty reactions in my
proteins of interest.

Anyone got any bright ideas as to what I might try ??


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