imidazole and degradation

Achim Recktenwald, PhD ARecktenwald at
Wed Jun 23 12:53:05 EST 1999

It's more likely you are co-purifying one or more proteases with your
Some of them are able to chop away at proteins even in 20 - 30% ammonium

The other possibility: I assume you are purifying a his-tagged protein. How
sure are you that the whole construct is expressed or that you do not have
mixtures of several expressed versions of different length of your protein.



Eva Horak <EM_Horak at> wrote in message
news:37700314.50CA9786 at
> Hi!
> We were wondering if anyone out there has experienced degradation of
> their his-tagged protein by high concentrations of imidazole (250 mM for
> example) when eluting from a nickel column?  Is there better way to
> elute proteins from these columns to avoid degradation?
> Thanks.
> -Lily

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