Western blotting was performed as described before [29 (link)]. Horseradish peroxidase (HRP)-conjugated 6×His-tag antibody (Abcam, Cambridge, UK) was diluted 1,000 times to detect the His-tagged proteins. The B. abortus monoclonal antibody (Thermo Fisher Scientific, Waltham, USA) or Y. enterocolitica O:9 monoclonal antibody (Fitzgerald, Acton, USA) was diluted 100 times to detect the polysaccharides of the glycoproteins. HRP-conjugated goat anti-mouse IgG (Abcam, Cambridge, UK) was used as the secondary antibody.
Protocol detail
Detection of His-tagged and Glycoproteins
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Variable analysis
independent variables
- Dilution of HRP-conjugated 6×His-tag antibody
- Dilution of B. abortus monoclonal antibody
- Dilution of Y. enterocolitica O:9 monoclonal antibody
- Detection of His-tagged proteins
- Detection of polysaccharides of the glycoproteins
- Western blotting protocol as described before [29]
- HRP-conjugated goat anti-mouse IgG as the secondary antibody
- Positive control: Not specified
- Negative control: Not specified
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