Immunological studies of the sperm-specific phosphoglycerate kinase-2 of mice

Antiserum was prepared to 500-fold purified PGK-2A from mouse testes. Depsite the apparent high purity of the immunizing antigen, double diffusion analyses showed two precipitin lines, one of which can be shown to correspond to PGK-2. An immunoabsorbent column of liver homogenate coupled to cyanogen bromid-activated Sepharose was used to remove the contaminating antibody. Both immuno-inactivation and double diffusion analyses demonstrated that the PGK-2 antigen appeared in testes between days 30 and 34 of postnatal development. The antisera prepared against highly purified PGK-2A did not disclose any immunological differences between PGK-2A, PGK-2B, and PGK-2C either by double diffusion analyses or quantitative immunoprecipitation. Antisera to PGK-2 did not inhibit in vitro fertilization of zona-free hamsters ova by mouse spermatozoa.