Figure 5.

Association of Act3p-His₆ with histone H2A during chromatography on a Ni²⁺-NTA column. Nuclear extracts prepared from strains MZ3 (A) and MZ3ACT3H (B) were applied to a Ni²⁺-NTA column and bound protein complex(es) were eluted with buffer containing increasing concentrations of imidazole (0.025–0.2 M). Aliquots of the applied extracts (AP), of the column flow through (FT), and of each fraction eluted from the columns were subjected to Western blot analyses with antiserum against Act3p (α-Act3p, upper panels) and anti-histone H2A monoclonal antibody (α-H2A, lower panels). Binding of the anti-histone H2A antibody to purified histone H2A is shown in B, right panel. Positions of Act3p and histone H2A on the blots are marked.