Dear all, I´m new in the field of crystallography. So please excuse my first question is off topic. I´m trying to purify a 182 kDa his-tagged bacterial enzyme. The expression in E. coli works quite well. After Ni-NTA the protein isn´t pure enough at all, so I tried several gelfiltration columns, but my enzyme interacts with the carbohydrate matrix, iex didn´t work either. So does anyone have suggestions? Perhaps a 182 kDa protein is too large to crystallize anyway?
Cheers, Marek
