Help! My HisTag Protein won't come off Ni Resin

Neil Saunders saunders at
Thu Feb 10 17:14:39 EST 2000

"Gareth" <gphillip at> wrote in message
news:38A2ED09.6922A38B at
> Pamela,
> Could be aggregated protein that's getting left behind. Do you c'fuge/
> your protein and have it in the same buffer as the column before loading?
> Or if you think the cysteines are causing a problem perhaps you could load
> the presence of DTT to keep them reduced.
> Hope that helps

But check the manual first-the concentration of DTT you can use is pretty
low, because it reacts with the nickel.  I think manufacturers generally
recommend beta-mercaptoethanol instead.

Neil Saunders

Department of Molecular Cell Physiology,
Faculty of Biology,
Vrije Universiteit,
De Boelelaan 1087,
1081HV, Amsterdam,
The Netherlands

tel: +31 20 4447194
fax: +31 20 4447229

email: saunders at

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