Figure 2

Blot overlay assays of 6His- Pf J with native 4.1R and GST-tagged 4.1R domains. The headings above each Western blot represent the proteins spotted on the nitrocellulose membrane. Glutathione-S-transferase (GST) and the relevant 4.1R domain served as negative and positive controls respectively. The membranes were overlaid with native 4.1R (4.1R overlay) and GST-tagged domains (10 kDa, 16 kDa and 30 kDa overlays) and binding detected with polyclonal rabbit anti-4.1R antibody using a chemiluminescent substrate. All the 4.1R fragments were detectable using the primary polyclonal antibody as demonstrated by the positive controls on the overlays. The 4.1R overlay indicates that 6His-Pf J binds native 4.1R. Two independent experiments (1 and 2) revealed specific interaction between 6His-Pf J and GST-10 kDa.