Fig. 8.

Effect of different amounts of SUMO protease on cleavage of SUMO-Nc-fusion. Serially diluted SUMO protease was added to 10 μg of purified SUMO-Nc-fusion protein and incubated in 30 μl PBS containing 5 mM β-mercaptoethanol at 30 °C for 1 h. The protease used was 20 U/μg. Aliquots (∼12 μl) from the incubation mixture were resolved on a 12% SDS–polyacrylamide gel and stained with Coomassie blue. Lanes: 0, uncleaved fusion (control); 1, 2 U of the protease; 2, 1 U; 3, 0.5 U; 4, 0.25 U; 5, 0.125 U; 6, 0.063 U; 7, 0.032 U; 8, 0.016 U; 9, 0.008 U; 10, 0.004 U.