Anti znf217 antibody

Thirty μg of protein were loaded onto 10 % SDS gel coupled with loading buffer. Then the protein was transferred to nitrocellulose (NC) membrane and the nonspecific binding sites were blocked using 5 % non-fat dry milk. Then the NC membrane was incubated with diluted anti-ZNF217 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) (1:200), anti-total CREB antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) (1:200), anti-aromatase antibody (Abcam, Cambridge, UK) (1:1000), anti-VEGF antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) (1:200) at 4 °C for overnight. After washing with TBST, membrane was incubated with diluted peroxidase-conjugated secondary antibodies for 1 h at room temperature. At last, the protein signals were detected using ECL western blotting substrate.

The ovaries of OHSS and control rats were fixed with 4 % paraformaldehyde and embedded in paraffin. Then 5 μm sections were prepared which followed by deparaffinization and rehydration through a graded ethanol series. The tissue sections were blocked using rabbit serum for 1 h at room temperature and incubated with anti-ZNF217 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) (1:100) overnight at 4 °C in a dark chamber. After being washed with PBS, the sections were incubated with secondary antibody (1:400) for 1 h at room temperature and then the color reaction was visualized by exposure to diaminobenzidine (DAB) for 2 min. To test the specificity of immunocytochemical staining, separate tissue sections were exposed to preimmune serum instead of the primary antibody (negative control).