FAQ: How do I separate MBP and TEV Protease from the protein of interest?

Both MBP and TEV Protease contain polyhistidine tags that can be used to remove them from the protein of interest. The TEV Protease digestion can be directly loaded onto NEBExpress Ni Spin Columns (NEB #S1427) or NEBExpress Ni Resin (NEB #S1428), since these resin formats are chemically resistant to the DTT in the TEV Protease reaction buffer. If using NEBExpress Ni-NTA Magnetic Beads (NEB #S1423) or another immobilized metal affinity chromatography format, we recommend dialyzing the TEV Protease digestion into a compatible buffer prior to IMAC.