Skip to main content
. 2018 Nov 7;7(4):39. doi: 10.3390/antib7040039

Figure 2.

Figure 2

Purificaiton of the zinc-transporter SsZntA from Shigella sonnei: (Reverse-affinity chromatography) R-IMAC. (a) Reverse (second) affinity chromatography profile: The first 60 mL represent the flow-through containing cleaved SsZntA (without HisTag); at 80 mL, corresponding to 250 mM imidazole, uncleaved SsZntA is eluted; while at 90 mL, at 500 mM imidazole, HisTagged TEVp is eluted. (b) On top, Coomassie stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and on the bottom, Western blot anti-HisTag; lane 1: Solubilized membranes, lane 2: Clarified solubilized membranes, lane 3: Affinity-chromatography purified SsZntA (with HisTag), lanes 4 to 6: Flow through of reverse affinity chromatography (SsZntA without HisTag) corresponding to retention volume from 0 mL to 60 mL, lanes 7 to 9: Eluted fractions corresponding to retention volume of 80, 85, and 90 mL, respectively.