Sc 515903

After deparaffinization of the sections, antigen retrieval was accomplished by boiling in citric acid–sodium citrate buffer (0.01 M, pH 6.0) for 15 min, and endogenous peroxidase blocking was performed using the same method as described above. Then, sections were incubated with a working solution of primary rabbit anti-vimentin monoclonal antibody (ZA-0511; Zhongshan Golden Bridge Biotechnology Co., Beijing, China) at 4 °C for 12 h. Next, sections were incubated with an anti-rabbit secondary antibody (ZF-0511, diluted 1:400; Zhongshan Golden Bridge Biotechnology Co., Beijing, China) at 37 °C for 1 h. Then sections were incubated with primary mouse monoclonal antibody against CYP27B1 (sc-515903, diluted 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, US) at 4 °C for another 12 h. The anti-mouse secondary antibody (sc-516141, diluted 1:50; Santa Cruz Biotechnology, CA, US) was added and incubated at 37 °C for 1 h, then nuclei were counterstained with DAPI (Neobioscience Biological Technology Co., Shenzhen, China), and sections were observed using immunofluorescence microscopy (Nikon, Tokyo, Japan).