Protein Separation

Achim Recktenwald, PhD achim at ibex.ca
Fri Nov 1 15:25:15 EST 1996


Bruce Amsden wrote:
> 
> "Brett S. Phinney" <brettsp at mail.utexas.edu> wrote:
> 
> >I need to separate and purify two very similar membrane proteins, and have
> >not been successful so far. They both have a very similar molecular weight
> >and presumably shape, because they only separate on an SDS-PAGE under
> >reducing conditions (unfortunately I need them non-reduced). They also seem
> >to have a similar hydophobicity plot, because I have not been able to
> >separate them using Reversed phase HPLC either. They do have differing
> >isoelectric points and have been separated using IEF in the past, but I
> >have not been able to figure out how to purify my proteins away from the
> >ampolytes.
> 
> >Does anyone have any suggestions on how I can separate and purify these?
> 
> >Any help would be greatly appreciated
> 
> >--
> >Brett Phinney
> >Cell Research Institute
> >University of Texas, Austin
> 
> Once you've purified your protein by IEF, can't you use gel filtration
> chromatography to get rid of the ampholytes?
> 
> Bruce Amsden


The latter usually never works. The stuff sticks to proteins like crazy
glue.

Achim



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