His-tag purification

[Mr. Chris Hoyle]

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
------------------------------------------------------