His tag specific polyclonal antibodies

For YebF samples induced 16–20 h, cells were pelleted, and the supernatant was harvested and precipitated with ice cold 10% trichloroacetic acid (TCA). For 4 h induction samples, culture volumes containing both cells and supernatant were harvested and directly TCA precipitated. For scFv13-R4, periplasmic fractions were harvested after spheroplasting cells in buffers containing 0.2 M Tris-Ac (pH 8.2), 0.25 M sucrose, 160 μg/ml lysozyme, and 0.25 mM EDTA. For RNaseA, protein was purified by nickel-affinity chromatography from the periplasmic fractions of 50–100 ml cultures. In all cases, protein was solubilized in Laemmli sample buffer and resolved on SDS-polyacrylamide gels (BioRad). Western blotting used 6x-His tag-specific polyclonal antibodies (Abcam) or C. jejuni heptasaccharide glycan-specific antiserum hR6 ^{29 (link)}. Pierce enhanced chemiluminescent (ECL) substrate (Thermo Scientific) was used for detection of bound antibodies. All blots were visualized using a Chemidoc^™ XRS+ system with Image Lab^™ image capture software (BioRad).