His-tag purification: imidazole quantitation?

Arle Kruckeberg arle at biovax.leeds.ac.uk
Fri May 19 04:52:07 EST 1995


Hi--

I too am purifying "my" protein by nickel-affinity chromatography of a
his-tagged recombinant...I'd like to know what the imidazole
concentration is in the fractions of the gradient, with the intention
of doing step elution. Does anyone have a method for quantifying the
free imidazole in the eluate, in the presence of all that protein?

Thanks!

Arle
*************************************************
a.k.a. Dr. Arthur L. Kruckeberg
 Department of Biochemistry and Molecular Biology
University of Leeds        phone  +44 +113 2333172
Leeds LS2 9JT              FAX    +44 +113 2333167
Great Britain              arle at biovax.leeds.ac.uk
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