Help! My HisTag Protein won't come off Ni Resin
saunders at bio.vu.nl
Thu Feb 10 17:14:39 EST 2000
"Gareth" <gphillip at peds.bsd.uchicago.edu> wrote in message
news:38A2ED09.6922A38B at peds.bsd.uchicago.edu...
> 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.
Department of Molecular Cell Physiology,
Faculty of Biology,
De Boelelaan 1087,
tel: +31 20 4447194
fax: +31 20 4447229
email: saunders at bio.vu.nl
More information about the Methods