Figure 3.

Optimization of reaction time of one-pot double labeling of TM₄₅₆AL via SML and SPAAC: TM₄₅₆AL (10 µM) was incubated with Gly₂-Bodipy (400 µM), SrtA (5 µM) and DIBO-AF647 (50 µM) in 20 mM Tris (pH 8.0), 150 mM NaCl and 5 mM CaCl₂ at 37 °C for different time (1–240 min): (A) SDS-PAGE followed by fluorescent scanning (Bodipy: Ex/Em=488/526 nm; AF647: Ex/Em = 633/670 nm) and (B) in-gel fluorescence intensities calculated and compared by Image J. The highest intensity of SML or SPAAC was set as 100%. Data points represent the mean ± SD from three independent experiments.