Figure 2 | Scientific Reports

Figure 2

From: Flavin Reductase Contributes to Pneumococcal Virulence by Protecting from Oxidative Stress and Mediating Adhesion and Elicits Protection Against Pneumococcal Challenge

Figure 2

Refolding of rFlaR. (a) rFlaR was eluted from a Ni-NTA column under denaturing conditions and dialyzed against a buffer containing 0.1% SDS. The solution was then dialyzed against: lane 1, 20 mM acetate pH 5.0; lane 2, 20 mM MES pH 6.0; lane 3, 20 mM phosphate pH 7.0; lane 4, 20 mM phosphate pH 8.0; lane 5, 20 mM Tris pH 9.0. All buffers contained 150 mM NaCl. Lane 6, rFlaR before dialysis. The dialyzed solution was resolved on 1D SDS-PAGE and stained with Coomassie Brilliant Blue. (b) rFlaR, solubilized with buffer containing 0.1% SDS, was dialyzed against PBS containing 2 mM of the following metal salts: lane 1, rFlaR before dialysis; lane 2, 2mM MnCl2, lane 3, 2mM  FeSO4, lane 4, 2mM CoCl2, lane 5, 2mM NiCl2, lane 6, 2mM CuSO4, lane 7, 2mM ZnCl2, lane 8, 2mM MgCl2 and 2mM CaCl2. The dialyzed solution was resolved on 1D SDS-PAGE and stained with Coomassie Brilliant Blue.

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