Figure 2.

Interaction between His-GlpF and HA-GlpF. Membranes from bacteria expressing His-GlpF, HA-GlpF, or both were extracted in 3% OG and incubated for 1 hr at 4°C. The solutions were then mixed with Ni-NTA magnetic beads and eluted with imidazole. (Note that only His-GlpF or proteins associated with His-GlpF will adsorb to Ni-NTA beads.) The eluates were analyzed by SDS/PAGE gels stained with Coomassie brilliant blue (CBB) or probed with monoclonal antibodies to His (α-His) or HA (α-HA). (A) Membranes from a bacterial culture coexpressing His-GlpF and HA-GlpF from a bicistronic operon were adsorbed to Ni-NTA and eluted. (B) Membranes from a bacterial culture expressing His-GlpF and from a second culture expressing HA-GlpF were mixed before adsorption to Ni-NTA and elution.