imidazole vs. histidine

Artem Evdokimov AEVDOKIMOZ at cinci.rr.com
Wed Jan 23 20:25:09 EST 2002


Imidazole worked for us just fine in a wast majority of cases. In one case,
addition of imidazole at a latter stage greatly improved the quality of the
crystals (upcoming paper in acta D). I have no evidence of detrimental
effects. One note - old or impure imidazole can contain stuff that's not
great for macromolecules. However, same applies to many other scientific
reagents :)

A.G.E.

P.S. You may or may not know about he curious effect of imidazole on
SDS-PAGE samples - namely that if you boil your samples with imidazole in
them you frequently get mysterious cleavage and even complete disappearance
of protein bands in extreme cases ! I do not know if histidine would
accomplish the same, however I see no reason as to why not...

"Nikolai Scherbak" <nikolai.scherbak at nat.oru.se> wrote in message
news:a0510120ab8745aa510f5@[130.243.107.82]...
> Dear collegues!
> I am working with purification of a recombinant His-tagged protein
> using Ni-NTA column. I am wondering if there is any difference in
> using imidazole vs. L-histidine for elution? Imidazole is cheaper
> but, as I heard, it can influence activity of my protein, so
> L-histidine is more recommended. Can it be so?
> Thanx in advance
>
> Nikolai
> --
> ************************
> Nikolai Scherbak
> Orebro University
> Fakultetsgatan 1
> S-70182, Orebro, SWEDEN
> tel.+46-19-301034
> fax +46-19-303566
>
> ---





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