Hi,
I am using Strep-Tactin® Superflow® high capacity resin by IBA to purify a Twin-Strep tagged fusion protein at N-terminal. I am facing issue to elute out the protein. For elution I have tried 2.5mM to 10mM desthiobiotin; however a large amount of protein is still stays on the beads. Any suggestion to solve this issue.
Thank you in advance
Hi Paras,
during your purification, are you working in batch or in a column? If you are working in batch, it can help to use a packed column where the resin bed is fixed with a frit. The directed flow of the elution buffer in a column helps to “wash down” the eluted protein to parts of the resin bed where the binding sites are already occupied, so the protein cannot bind again.
Otherwise, you can also elute your protein from Strep-Tactin® Superflow® high capacity resin via a pH shift. The bond between the tagged protein and the resin is stable at pH 7-8, anything above or below will lead to a dissociation of the resin.
If you have any further questions, I am happy to help.
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