HIS re-purification

Duncan Clark junk at hgmp.mrc.ac.uk
Fri Jan 23 11:48:46 EST 2004

Historians believe that in newspost <bur0r9$1ckg$1 at gwdu112.gwdg.de> on 
Fri, 23 Jan 2004, Stephan Wenkel <wenkel at mpiz-koeln.mpg.de> penned the 
following literary masterpiece:
>Thanks for your reply, but stripping the resin with EDTA  does also have
>only a little effect.

Then it's not binding by His-Ni interaction, as EDTA removes all the Ni 
of the resin.

Hydrophobic interaction? Try 20% Ethylene glycol in buffer.

Try 20% ethylene glycol plus a Urea gradient up to say 3M in buffer.

Or wild guess.

Could it be binding to the chelating bit of the column i.e. You 
initially purify your protein which hangs onto some Ni. You load your 
dialysed protein back on and create a chelated protein via the Ni 
moiety, with a very strong interaction.

The difference between theory and reality is much smaller in theory
then it is in reality

Duncan Clark
GeneSys Ltd.

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