Fig. 6
Final gel of Nickel purified recombinant zymogen (no leader) proteases grown and expressed at either 23 °C or 10 °C. In order to demonstrate that inactive zymogen proteases (with intact N-terminal his6-tag) can be isolated, AaSPVII grown at 23 °C (a) and AaLT grown at 10 °C (b) were purified. These samples are the post-dialysis concentrate of AaSPVI-NL and AaLT-NL dialyzed in 50 mM Sodium Acetate pH 5.2 + 2 mM DTT (buffer exchanged twice and set at 4 °C). Samples loaded on the gel are in micrograms (μg) and are increasing in order to show that the single step purification scheme led to a near homogenous sample with very little contaminants