This approach should work for a His-tagged protein. The protein will bind to the column and the labels will flow through.

We have done this before but be careful of the amount of DTT you add to quench the maleimide before adding it to the column and be sure to extensively wash before eluting your labeled protein

Thanks Adam and David. It will help me a lot.

Another doubt: Is there any preference for the washing buffer? Does it need to have imidazole as an usual purification procedure?

The protein has presumably already been purified, so there is no need to include a low concentration of imidazole in the binding buffer to prevent non-specific protein binding.

Hi everyone,

I have applied this approach in the last week and it indeed worked very well. The only detail is that I used a buffer with 20 mM of imidazole as wash buffer because it was what I had in hands at the time.

Thank you very much for your help!

Best regards,

Gustavo