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. 2009 Sep 10;37(20):6754–6764. doi: 10.1093/nar/gkp748

Figure 2.

Figure 2.

Mapping of Rad51 interaction by in vitro pull-down assay. Purified Srs2, srs2 1–998, srs2 1–910, srs2 1–890, srs2 1–875 and srs2 1–860 were mixed with Affi-Rad51 beads (A) or Affi-BSA beads (B) The supernatant that contained unbound protein (S), wash (W) and the SDS eluate (E) were resolved by SDS-PAGE and stained with Coomassie Blue. (C) In panel I, Purified srs2 Δ875–902, 1 µg, analyzed by SDS-PAGE and stained with Coomassie Blue. In panel II, Srs2 and srs2 Δ875–902 were combined with purified Rad51 and mixed with nickel NTA agarose beads. The supernatant that contained unbound protein (S), wash (W) and the SDS eluate (E) were resolved by SDS-PAGE and stained with Coomassie Blue (lanes 1–6). As control, Rad51 alone was incubated with nickel NTA agarose beads (lanes 7–9). In panel III, Srs2 and srs2 Δ875–902 were examined for PCNA interaction (lanes 1–9) following the same procedure in panel II.