Monoclonal anti his tag

Recombinant proteins rHis-SseB and rGST-SseB were analyzed by western blotting using the monoclonal anti-His tag (Sigma, Saint Louis, MO, USA), GST mouse mAb (Cell Signaling Technology, Beverly, MA, USA), or antisera against Salmonella Enteritidis C50041 prepared by orally infecting mice with Salmonella Enteritidis C50041. Western blotting was performed as described previously [19 (link)]. Briefly, the contents of the recombinant proteins were separated by SDS-PAGE and electrotransferred to a nitrocellulose membrane. The membranes were probed with a monoclonal anti-His tag (1:3000), anti-GST tag mouse mAb (1:3000), or antisera against Salmonella Enteritidis C50041 (1: 1000) as the primary antibody and then incubated with horseradish peroxidase (HRP)-conjugated goat anti-mouse immunoglobulin G (IgG) antibody (1:5000 dilution; Sigma). Immunoreactivity was measured using Super Signal West Pico Chemiluminescent Substrate (Pierce, Chemical, Rockford, IL, USA) according to the manufacturer’s instructions.