Dear All, My protein is cloned in pET-15b vector, contains His- tag, thrombin cleavage site and extra sequence of 20 amino acids from vector. I crystallized without cleaving extra sequence and never got any crystal hits/crystals. Once, I tried thrombin reaction and it didn't work. Here I would like to know how efficient the cleavage by thrombin and it is kind of you, can provide protocol and buffer system for reaction.
Thank you Venkat.
