Affinity purification of a liver protein

[rdudley at aherf.edu]

In article <3598FFD4.245D at animal.ufl.edu>,
  mgreen at animal.ufl.edu wrote:
>
> Hello,
> 	We are preparing to isolate a cytosolic liver protein using affinity
> chromatography.  We are not sure what the degree of expression will be,
> however it should be high.  My question is should any preliminary
> "clean-up" be performed on the cytosols prior to affinity purification.
> Perhaps a size exclusion or ion-exchange column?  Are there any
> standardized protocols for liver tissues?
>
> Thanks,
> Mike Green

Mike--

Very rarely are there standardized protocols for proteins.  Size exclusion,
ion-exchange, or ammonium sulfate are usual possibilities, but it all depends
on your protein.  There's always the chance you'll destroy your protein in
one of these steps, or they may be a panacea.  I'd try the affinity column
first, and wash it very very well.  If that's not satisfactory, try adding a
pre-purification step.	Good luck!

rich

Richard J. Dudley
Senior Research Assistant
Center for Genomic Sciences
Allegheny University of the Health Sciences
rdudley at aherf.edu


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