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

[Neil Saunders]

"Gareth" <gphillip at peds.bsd.uchicago.edu> wrote in message
news:38A2ED09.6922A38B at peds.bsd.uchicago.edu...
> Pamela,
>
> Could be aggregated protein that's getting left behind. Do you c'fuge/
filter
> 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
in
> 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

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