his-tagged protein purification with metal chelate columns

Tara McCready tara_mccready at darwin.biochem.ualberta.ca
Mon Jul 31 17:47:19 EST 1995


Are people generally using pre-charged nickel-NTA resin (ie from Qiagen)? 
What advantages does this have over iminodiacetic acid agarose (ie from
Pharmacia), other than the obvious need to charge it with nickel
yourself?   Some of the literature seems to suggest that zinc-IDA works as
well as Ni-NTA.....
I would appreciate any info because my his-tagged protein isn't sticking
to Ni-IDA nearly as well as I have seen reported (50mM instead of up to
300mM imidazole at pH 7.5 in Tris-HCl, 10 mM betaME, 5% glycerol).

Thanks,

Tara McCready
Dept Biochemistry
University of Alberta
tara_mccready at darwin.biochem.ualberta.ca



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