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. 2005 Sep 26;391(Pt 1):135–142. doi: 10.1042/BJ20050640

Figure 6. In vitro pull-down assay to demonstrate the self-binding ability of subregion IV.

Figure 6

(A) GST–rP[IV]–HA (500 ng) was incubated with 10 μl of glutathione–Sepharose beads, and the bead-bound protein was visualized by SDS/PAGE followed by silver staining. (B) Left-hand panel: 12.5 ng of the indicated proteins were visualized by anti-MBP Western blotting. Middle panel: the glutathione–Sepharose beads carrying GST–rP[IV]–HA and 400 ng of the MBP-tagged proteins were subjected to the in vitro pull-down assay as described in the Experimental section, and the pulled-down proteins were visualized by anti-MBP Western blotting. Right-hand panel: a similar experiment was performed as that shown in the middle panel, except for the absence of GST–rP[IV]–HA. Molecular-mass sizes are given in kDa.