Response to the comments made by Marcellus Ubbink [post]

Gottfried Otting
2020 unpublished
We are grateful for the insightful comments and identifying errors. Line 100: A 100-fold excess of TEV was used, that seems an awful lot for an enzyme. How was it removed? Was second NTA column used? Response: We appreciate detection of this error! TEV protease was added in 0.1 molar ratio to remove the His6 tag. In addition, a second 5 mL Ni-NTA column was used to remove the TEV protease from the protein. We propose to write in the revised manuscript: "the protein was dialysed into TEV protease buffer (50 mM Tris-HCl, pH C1
doi:10.5194/mr-2020-26-ac1 fatcat:wfgpvmu34nbnppivezhjqnbjuy