Dear all, I am trying to express the human protein using bacterial expression strain (Rosetta) and purified using Ni-NTA affinity purification. The Molecular weight of out protein is 47 kDa. In SDS-PAGE, we have seen that 27 kDa contaminant protein co-eluted with our protein even at high concentration of Imidazole. In Superose 12 column, these two proteins eluted as a single peak and its corresponding molecular weight suggestive of partial interaction. By mass mapping we have found 27 kDa band is an adenylate kinase. Is there any specific way to separate adenylate kinase from our protein? Thanks in advance,
Sivaraman Padavattan
