FIGURE 10.

AntitoxP binds directly to ToxP. Cell‐free IVTT reactions containing GST‐tagged AntitoxP (GST‐A27), GST‐A27 and V5‐tagged ToxP (A28‐V5), A28‐V5 or GST‐A27, and XpressT‐tagged CBU00665 (XpressT‐665) (Figure S9) were incubated with GST‐affinity beads. After incubation beads were washed and bead‐bound proteins were eluted using a reduced glutathione buffer. Flow‐through (F), elution 1 (E1), and elution 2 (E2) fractions were analyzed by immunoblot probed simultaneously with (a) anti‐XpressT and anti‐V5 or (b) anti‐GST. GST‐AntitoxP pulldown of only ToxP‐V5 was observed