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Dear Fred -- just to check... are you sure you have the His-tag? Might have been cleaved somehow? You might want to increase the number of His in the tag as well. HTH. -- Leo -- On 27 Jan 2009, at 21:00, Fred wrote: > Hi ccp4 list, > I am trying to purify a his-tag protein by metal affinity > chromatography. The protein was expressed in inclusion bodies and > its his-tag doesn't bind the Qiagen Ni resin in denatured > conditions (urea 8M or GndHCl 6M). Playing with NaCl and detergents > didn't help much. > Any help is appreciated. > Fred ------------------------------------------------------------ Chavas Leonard, Ph.D. @ home Research Associate Marie Curie Actions Fellow ------------------------------------------------------------ Faculty of Life Sciences The University of Manchester The Michael Smith Building Oxford Road Manchester Lancashire M13 9PT ------------------------------------------------------------ Tel: +44(0)161-275-1586 e-mail: [log in to unmask] http://personalpages.manchester.ac.uk/staff/leonard.chavas/