Figure 4.

Native PAGE and qualitative SEC analyses to compare the stability of the in vivo-associated and on-column-associated complexes of Scc4:His₆-Scc1. (A) Native PAGE gel of the in vivo-associated complex (lane 1) and the on-column-associated complex after purification (lane 2) and 2 days at room temperature (lane 3). (Native PAGE analysis of the in vivo-associated complex after 2 days at room temperature is shown in Figure 6, lane 2 from a second stability study.) Lane 4 is the Fisher BioReagents EZ-Run Rec pre-stained protein ladder with molecular weights listed to the right in kDa; the ladder was included as a qualitative guide. The full gels from which these lanes were extracted are shown in Figure S4. (B,C) Qualitative SEC analysis to assess the stability of (B) the in vivo-associated complex after purification (0 d, black solid line) and after 2 days at room temperature (dashed black line) and (C) the on-column-associated complex after purification (0 d, gray solid line) and after 2 days at room temperature (dashed gray line). Differences in the elution times between the (B) in vivo- and (C) on-column-associated complexes are not interpretable (refer to Section 2.5 for details).