Dear CCP4bb users (this is an off-topic question), We have a few proteins being expressed as HIS-TAG_(TEV_cleavage_site)_PROTEIN and we are about to initiate the purification steps.
We have already used the HiTrap-Chelating columns from GE for the first purification step (affinity chromatography) and we would like to move forward with TEV digestion and a second purification step. We know these following steps are very protein-dependent, but we were wondering one could share his/her experience in the following steps: removal of imidazole, cleavage protocol and cleavage identification, second chromatography, etc. Any experience would be appreciated. Thanks in advance Ronaldo.
