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. Author manuscript; available in PMC: 2014 Feb 20.
Published in final edited form as: Chem Commun (Camb). 2009 Oct 12;46(1):76–78. doi: 10.1039/b915612g

Fig. 1.

Fig. 1

NuPAGE analysis of cysteine-N3 ligation by intein-mediated cleavage. Luc8-pep-GyrA protein (lane 1) was incubated with different thiol groups for 16 h at 4 °C: 10 mM Cys-N3 (lane 2), 20 mM MESA (lane 3), and 20 mM MESA/10 mM Cys-N3 (lane 4). The purified sample from lane 4 is displayed in lane 5. The reaction mixture was loaded into each well after mixing with lithium dodecyl sulfate (LDS) loading buffer in the absence of reducing agent. The arrows indicate the position of Luc8 protein fused with the Cys-N3. The molecular weight standard is displayed in the far left lane. Electrophoresis was carried out on 12% gel in bis (2-hydroxyethyl)amino-tris(hydroxymethyl)methane (Bis-Tris) buffer containing LDS for 45 min.