Figure 9

Effect of salt concentration, temperature and EDTA on p35 gel shift assay. Gel retardation assay was performed either using no extra salt or different concentration (lanes 1–5) of NaCl. Band shift was completely inhibited at 0.5 M NaCl concentration and above. Effect of temperature on binding reaction was tested by incubating the reaction mixture (lanes 6–9) at different temperature. Binding reaction was completely inhibited at 55°C and above. All the other reactions were carried out at room temperature. Effect of bivalent cations were tested by incubating the reaction mixture in the presence of 50 mM and 100 mM EDTA (lanes 10–11). Presence of EDTA completely inhibited the binding reaction.