protein electroelution from gels, does it modify the protein?

Federici federici at icgm.cochin.inserm.fr
Wed Apr 23 09:23:15 EST 1997


In article (Dans l'article) <335648F2.3686 at bris.ac.uk>,
roger.starr at bristol.ac.uk (Roger Starr) wrote (écrivait) :

> Trying to purify myosin heavy chain,have tried column filtration but 
> seperation not good enough. Thought of running sds gels and cutting out
> the protein and then electroeluting but boss seems to think this may 
> alter the protein in some way. Anybody had any experince of this?

When electroelution is used to recuperate protein from gel, i have never
had problems. First of all electroelution on hydroxyapatite using
electrophoresis on tubes. the second step will be washing of
hydroxyapatite with 1mM sodium phosphate pH 6,8, 0,1% SDS, 1mM DTT; the
procedure will be ended carrying elution from hydroxyapatite using a
higher concentration of the buffer phosphate (0,7M) or better a gradient
20mM-0,7M in order to obtain a well-purified protein.



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