Objective: To prepare anti-mouse ZP2 (mZP2) antibody and study the immune activity of antiserum.
Methods: The mZP2 recombinant protein was induced and expressed by Rosetta host bacteria, the recombinant ZP2 protein was separated by electrophoresis, and the immunized antigen was prepared by cutting gel; the emulsified antigen was used to actively immunize New Zealand male rabbits to prepare antiserum; the antibody response level was determined by ELISA; the antiserum was determined by immunohistochemistry Specificity; the effect of antiserum on mouse egg-binding acrosome reaction was detected by sperm-egg binding assay.
RESULTS: The results of ELISA showed that antiserum was induced by immunizing New Zealand male rabbits with recombinant protein. Immunohistochemical experiments showed that the antiserum reacted specifically with the mouse ovary zona pellucida. The sperm-egg binding experiments showed that the anti-ZP2 antibody could inhibit the sperm-egg binding after acrosome reaction.
Conclusion: The recombinant mZP2 protein induces the production of anti-ZP2 antibody, and the antibody can inhibit the combination of sperm and egg after acrosome reaction.