histag elution alternative

Frank Fuerst ffrank at rz.uni-potsdam.de
Thu Nov 2 10:27:58 EST 2000


"Richard P. Grant" <rpg14 at yahoo.co.uk.invalid> wrote:

>In article <8tq8il$8nq$1 at news.doit.wisc.edu>, 
>klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu (Dima Klenchin) wrote:
>
>> The nickel is gone. We are talking about _recharging_ and that 
>> necessarily implies stripping the original, fully or partially reduced,
>
>From Cornelius' posting, that's not that crystal clear.   I'm not sure 
>whether he reduced it in the nickel-bound state or not.  

I would even say that it _is_ clear that the nickel was still there.
The original poster wanted an alternative for imidazole as an eluent,
and asked wether EDTA or DTT would work:

>We now want to turn to an alternative elution 
>approach e.g. EDTA. [...]
>
>I have also been thinking about whether one could 
>use high concentrations of DTT. In the manual 
>(Quiaexpressionist) they write that DTT can reduce
>Nickel thereby preventing nickel from binding His-tagged
>proteins.

And Cornelius warned him to use DTT.

Bye, Frank
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