His-tag purification

Mr. Chris Hoyle bmbckh at biovax.leeds.ac.uk
Thu Aug 7 08:57:20 EST 1997

We use Ni-NTA agarose from Qiagen for purification of H6-tagged
proteins which are subsequently used for CD crystallisation studies.
Some people batch bind whilst I prefer binding on the column overnight
by recirculation at 0.5ml/min. We bind in the presence of 20mM
imidazole, 50mM Na phos. (pH 8.0), 10% glycerol and 0.1%
dodecylmaltoside (our proteins are integral membrane proteins). We have
never needed to try other resins because the resin from Qiagen performs
so well. 

Chris Hoyle

Department of Biochemistry and Molecular Biology
University of Leeds             phone +44 0113 2333172
Leeds LS2 9JT                   FAX   +44 0113 2333167
Great Britain           e-mail BMBCKH at biovax.leeds.ac.uk

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