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. 2011 Jul 5;5(7):e1225. doi: 10.1371/journal.pntd.0001225

Figure 5. Determination of rTs-Pmy binding to human C8 and C9.

Figure 5

Human C8 (1 µg, Lane 1) and C9 (1 µg, Lane 3) and non-relevant BSA control (1 µg, Lane 2) were subjected to SDS-PAGE under reducing conditions and (A) stained with Coomassie blue, and then (B) transferred onto a PVDF membrane, incubated with rTs-Pmy (5 µg/mL) and detected with anti-rTs-Pmy mAb (50 ng/mL). The same materials that were added, plus non-relevant control protein rTs87 (300 ng; Lane 4) were subjected to SDS-PAGE and (C) stained with Coomassie blue, and (D) transferred to an NC membrane, incubated with rTs87 (5 µg/mL) and detected with anti-Ts87 rabbit serum (1∶4000). To determine the binding competition between soluble and immobilized C9 to rTs-Pmy, rTs-Pmy (5 µg/mL) was pre-incubated with C9 (30 µg/mL) for 1 hour before incubation with the membrane containing with C9 (1 µg) and BSA (1 µg), and was detected with anti-rTs-Pmy mAb (50 ng/mL) (E). M:standard protein marker.