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.
Duncan
--
The difference between theory and reality is much smaller in theory
then it is in reality
Duncan Clark
GeneSys Ltd.
