imidazole and degradation

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


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

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.

Cheers,

Achim


Eva Horak <EM_Horak at fccc.edu> wrote in message
news:37700314.50CA9786 at fccc.edu...
> 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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