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

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

In article <39AFAFA5.F23D687C at>, 
silke.meister at (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 

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 

Richard P. Grant MAD Phil
Structural Studies    
MRC-LMB                         Please reply to rpg 'at'

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