getting rid of lipids

J. Norrie 4104jnor at vm1.ulaval.ca
Wed May 24 10:18:32 EST 1995


> Does anyone have any trick for getting rid of lipids from a total protein
> extract? I'm purifying proteins from plant extracts and the lipids 
really gunk
> up the ion-exchange clomns that i use in the first two steps of the
> purification.  So far my only recourse has been to wash the columns alot with
> NaOH, and to pour new ones often, but this is rather expensive (and a pain).
> I'm looking for any method that would work for either a small scale (0.5 ml
> sample) or large scale (1l)protein prep.
>I also had the same problem..
Solution: 
1) Treat your leaves with 100% cold acetone. Wash the leaves several times
with Acetone, this might help in removing pigments and phenolics.
2) Air dry to remove all residual acetone(takes about 20-30 min.depending
on the amount of starting material)
3) treat these dried leaves with n-hexane.
4) air dry, or else after hexane treatment you could give one wash of
acetone and then air dry
5) grind the tissue (normally you don't require liq.nitrogen. since tissue
is dry) and add 10 vol of extraction buffer per gm dry weight. Incubate at
room temperature for 30 min.And then spin at 14000 rpm to get proteins in
supernatent. This protocol takes care of both phenolics and lipids. You
may additionally include in the extraction buffer, if you want,
antioxidants. 
Savita



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