Hrp labeled goat anti rabbit antibody

The slides were heated at 60°C for 2 hours, deparaffinized in xylene, and rehydrated by graded ethanol. After antigen retrieval, the tissue sections were blocked with goat serum. The sections were then exposed to antibodies against δ‐catenin or Ki67 (Abcam, ab15580) overnight at 4°C. The slides were washed with PBS and incubated with HRP‐labeled goat anti‐rabbit antibody (ThermoFisher, #31460) for 1 hour. After washing, the slides were added with diaminobenzidine (DAB) and counterstained with hematoxylin. In the end, the sections were dehydrated by graded ethanol, sealed by neutral balsam, and visualized with microscope (at 400 × magnification).

The ovarian tissue slices as above mentioned was utilized to immunohistochemistry examination. Sections were incubated with primary antibodies against α-SMA (55135-1-AP, Proteintech, Wuhan, China) or CD31 (A11525, Abclonal, Wuhan, China) overnight at 4°C. After washing in PBS, slides stained with α-SMA were incubated with goat against rabbit biotin-labeled antibody (A0277, Beyotime, Shanghai, China) in PBS solution for 60 min at 37°C and conjugated with HRP-labeled Streptavidin (A0303, Beyotime) for 30 min at room temperature. The HRP-labeled goat anti-rabbit antibody (#31460, Thermo Fisher Scientific) was prepared to conjugate with CD31. Then slides were colorated using DAB reagent and counterstained with hematoxylin. The images were shot at × 100 or ×400 magnification.