Abstract: Objective To prepare rabbit anti-mouse zona pellucida 2 (mZP2) polyclonal antibodies and test their
immunoactivity. Methods Recombinant proteins of mZP2 expressed in Rosetta transformant was separated by SDS-PAGE, and
the gel strips containing the recombinant mZP2 were cut out and emulsified to immunize New Zealand white rabbits. The
antibody response of the antiserum was detected by ELISA, and the specificity of the antiserum was verified by
immunohistochemical assay. The effect of the antiserum on the binding of oocytes with acrosomal reacted sperm was tested by
sperm-egg binding assay. Results ELISA results showed that the immunized rabbit produced anti-mZP2 antiserum. The
antiserum reacted specifically with the zona pellucida of mouse ovarian sections. Sperm-egg binding assay showed that
treatment of the oocytes with the anti-mZP2 antiserum caused decreased binding of zona pellucida with the acrosomal reacted
sperm by 43.7%. Conclusion We obtained rabbit anti-mouse ZP2 polyclonal antibodies that can inhibit the binding of oocytes
with acrosomal reacted sperm.