Skip to main content
. 2016 Apr 25;113(19):5406–5411. doi: 10.1073/pnas.1600546113

Fig. 2.

Fig. 2.

Functional analysis of r-0583 and r-0582. (A) Pull-down interaction assay between r-0583 and naïve goat serum. r-0583 was bound to a resin and incubated with serum. After extensive washing, bound proteins were eluted and analyzed by SDS/PAGE in reducing (Left) and nonreducing conditions (Right). Control experiments included the bait protein r-0583 without serum (lane: r-0583) and the naked resin incubated with the serum (lane: serum). Black arrow, IgG chains (55 kDa, heavy; 25 kDa, light; ∼250 kDa, whole IgG). (B) Effect of r-0582 on IgG pulled down from goat serum. Proteins eluted from the pull-down experiments were mixed with purified r-0582 (lane: pull down + r-0582) and incubated for 15 min at 30 °C. Samples were checked by SDS/PAGE in reducing (Left) and nonreducing conditions (Right). Black star, cleaved IgG heavy chain. (C) Cleavage of purified goat IgG by r-0582. Samples were checked by SDS/PAGE and the IgG fragments were detected by Western blot. Nonspecific detection of r-0583 was caused by its nonimmunological binding of the primary antibody.