B abortus monoclonal antibody

IPTG-induced cells (1 mL) were collected by centrifugation and then resuspended in 100 µL of ddH₂O. The whole-cell or purified samples were mixed with 1:1 (v/v) 1× Loading Buffer (100 mM Tris-HCl, pH 6.8, 3.2% (w/v) SDS, 0.04% (w/v) bromophenol blue, 16% (v/v) glycerol, and 40 mM DL-dithiothreitol). The samples were placed in a boiling water bath for 10 min and then cooled to room temperature. After centrifugation at 10,000 ×g for 10 min, 10 μL of each sample was subjected to SDS-PAGE and western blotting using the methods described in our previous work [38 (link)]. Horseradish peroxidase-conjugated 6×His-tag antibody (Abmart, Shanghai, China) was used to detect the His-tagged proteins. A B. abortus monoclonal antibody (Thermo Fisher Scientific, Waltham, MA, USA) was diluted 1:100 and used to detect polysaccharides of B. abortus in the glycoproteins. A Y. enterocolitica O:9 monoclonal antibody (Fitzgerald, Acton, MA, USA) was diluted 1:200 and used to detect Y. enterocolitica O:9 polysaccharide in the glycoproteins.