gst elution

Robert Solomon Bioc rgs at mole.bio.cam.ac.uk
Thu Nov 24 05:46:33 EST 1994


In article <Czq5Iw.FoG at phibred.com> bipin dalmia <dalmiabk at phibred.com> writes:
<snip>
>
>do you or anyone else have any data on GST binding to GSH-agarose (or
>sepharose) in the presence of urea? i am trying to use some urea (1-2 M)
>in my lysis buffer to get more of my gst fusion soluble.
>
 Well, I've had success working at 2M urea, getting a GST-fusion protein on
and off beads and cutting successfully with (human) thrombin.  I had to 
break up inclusion bodies at 8M urea, dilute to 2M, and carry on as usual.
Of course, the question of solubility of your target protein then becomes
the main problem.

Good luck

Rob Solomon
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