Western Blot Analysis of T. gondii Immunogenicity

This immunogenicity analysis referred to the previous study (21 (link)). The rTgBFD1 was separated by 12% SDS-PAGE gel and transferred to the nitrocellulose membranes (Millipore, Shanghai, China). The blots were blocked with 5% skim milk in phosphate-buffered saline (PBS) with 0.5% Tween 20 (PBST) for 1 h at 37°C and then incubated with the serum collected from normal or T. gondii-infected mice (1:100 dilution) for 1 h at 37°C. After three washes, 5 min each using PBST buffer, the blots were incubated with goat anti-mouse IgG-HRP antibody (Sigma, Shanghai, China) (1:5000 dilution) for 1 h at 37°C. After three washes, 5 min each using PBST buffer, the blots were finally visualized by the enhanced chemiluminescence kit (Vazyme, Nanjing, China).