FIG. 6.

Affinity purification of His₆HigB and associated HigA-Myc. His₆HigB and HigA-Myc were coexpressed from pQE30HigBAmyc in E. coli XL1Blue, and His₆HigB was affinity purified from cell extracts, using Ni-NTA resin under nondenaturing conditions. Cell extracts and subsequent fractions from the purification were run in denaturing polyacrylamide gels and then analyzed using either Coomassie blue staining or Western blotting. (A) Proteins from various purification stages stained with colloidal Coomassie blue. (B) Western blot of pre- and postpurification fractions probed with anti-Myc antibody. (C) Western blot of pre- and postpurification fractions probed with anti-His₆ antibody.