On Thu, 2011-09-15 at 22:20 -0400, m zhang wrote:
> Second is about reusing of Ni-NTA resin. According to Qiagen's
> instruction, after using fresh Ni-NTA resin, one only needs to wash
> the used Ni resin first with 0.5M NaOH, then with your own buffer.
> After that the resin is ready to be reused until it needs being
> recharged. But my question is: Once immidazole competes with
> His-tagged protein and binds to Ni-resin, how can immidazole be rinsed
> off with the same buffer(usually pH is above 7) one uses to purify the
> protein?
Imidazole binds much weaker than a his-tag, and thus more of it goes
into the buffer when you wash the column. In theory, if you wash a
column with protein bound long enough, it will all slowly come off.
--
"Hurry up before we all come back to our senses!"
Julian, King of Lemurs
