Figure 1: Purification and verification of His6-tagged intact VanS. | Scientific Reports

Figure 1: Purification and verification of His6-tagged intact VanS.

From: Hydrodynamics of the VanA-type VanS histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin

Figure 1

(A) SDS-PAGE and (B) Western blotting of mixed membrane samples of E. coli host cells either uninduced (U) or induced (I) with 1 mM IPTG. (C) and (D) SDS-PAGE and Western blotting (respectively) of purified intact VanS protein ((C) 10 μg in SDS-PAGE; (D) 1 and 5 μg in Western blotting). SDS-PAGE employed 4% stacking and 12% resolving gels and proteins were visualised using Coomassie blue staining. Western blotting detected the presence of the C-terminal hexa-histidine tag. Arrows denote the positions of monomeric VanS. (C) and (D) indicate the presence of multimeric VanS, even under denaturing conditions, as commonly observed during preparation of membrane proteins for SDS-PAGE (see Methods). M, molar mass markers.

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