Figure 7
Determination of optimal GuHCl concentration for analyte elution. 5′-biotinylated AR aptamer bound to streptavidin agarose resin was incubated with GST fused with the indicated tag. After washing, the bound protein was eluted using Tris-HCl buffer supplemented with the appropriate GuHCl concentration. Next, the protein samples were denatured and one fortieth of the volume of each sample was separated on 12% SDS-PAGE followed by Coomassie staining and analyzed using Multispectral Imaging System IMAGER with Launch VisionWorksLS. The results were normalized relative to signal from the protein sample eluted with buffer supplemented with 0 mM GuHCl (0% eluted protein). Recombinant GST protein tagged with: (A) R6, (B) R8 and (C) Tat49–57 peptide. Each panel presents the results of one of three independent experiments which is representative.
