GST-fusion protein doesn't bind to sepharose-beads

Richard P. Grant rgrant at netscape.net
Fri Sep 1 08:51:37 EST 2000


In article <39AFAFA5.F23D687C at molmed.uni-erlangen.de>, 
silke.meister at molmed.uni-erlangen.de (Silke Meister) wrote:

> I cloned my protein in the pGEX-vector and wanted to purify this
> GST-fusion protein, but I wasn't succesful!
> The protein didn't bind to the sepharose-beads. In my opinion the
> protein prevent the binding from GST to GSH, because the empty
> GST-vector without the protein can bind!
> I hope you can help me to solve my problem!

Silly questions, i know, but it'd help if you could confirm the 
following:

1) Did your fusion protein express?
2) If so, did it appear in the soluble fraction?
3) If so, did you see it in the flow-through from the seph beads, or 
what?

-- 
Richard P. Grant MAD Phil       http://www2.mrc-lmb.cam.ac.uk/personal/rpg/
Structural Studies              http://www.scienceboard.net/
MRC-LMB                         Please reply to rpg 'at' mrc-lmb.cam.ac.uk
                                






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