29 September 2015 15 5K Report

Hi all,

Most of my His-tagged protein does not bind to Ni beads comes out in flow through in the presence of 8 M urea. Please see the attached gel image to compare protein in supernatant (before loading to beads) and in flow through (after 45 min incubation with Ni-NTA). Shouldn't be binding efficiency increased as His residues were exposed? 

Thanks in advance,

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