Are reducing agents/denaturants compatible with the Proteus columns?
Reducing agents can reduce the resin matrix and adversely affect binding of the His-tagged protein to the Spin Column. Its inclusion depends upon whether the His-tagged protein elutes with contaminants as β-mercaptoethanol can reduce all disulphide bonds formed between the contaminating proteins and the target protein. We recommend 0.5 mM TCEP. TCEP is a more stable reducing agent and it is less likely to reduce free Ni2+. Do not use strong reducing agents such as DTT or DTE as these tend to reduce the metal ion, which will lower the binding efficiency of the IMAC column. Denaturants/solubilizing agents such as 8 M urea or 6 M guanidine hydrochloride are fully compatible with the columns.
