bacterial expression/protein purification

R. John Lye rjl6n at virginia.edu
Thu Apr 6 12:50:48 EST 2000


Ruth McMahon wrote:
> I am working with the pBAD bacterial his tag expression system from
> Invitrogen, and am trying to purify the his-tag protein. I have checked
> a number of methods to lyse the cells to release the protein, but in
> each case the vast majority of my protein stays in the pellet. 

Do you know that your recombinant protein is not going into inclusion
bodies?  They are quite resistant to most lysis methods and will require
harsh treatment to solubilize - on the other hand, the inclusion bodies 
can be used as a first round purification step.
-- 
John Lye
rjl6n at Virginia.edu




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