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兔附睾尾液经ＨＰＬＣ纯化后，获得分子量为２０ｋＤ的附睾特异蛋白，命名为ＢＥ－２０。ＢＥ－２０的抗体具有明显抑制人精子穿透去透明带仓鼠卵的能力。已获得编码ＢＥ－２０的０．５ｋｂｃＤＮＡ片段，命名为ＲＥＤ－２０。用地高辛标记此片段，与兔睾丸、附睾、输精管和肝组织进行原位杂交，观察ＢＥ－２０在各种组织中的定位和表达。采用杂交前热变性与不用热变性组织两种方法进行原位杂交。杂交信号均为紫褐色的颗粒，分布在附睾尾的主细胞和近端输精管的上皮细胞的胞质内，表明胞质内ｍＲＮＡ与地高辛标记ＲＥＤ－２０ｃＤＮＡ探针杂交。组织经热变性后，附睾尾和近端输精管上皮细胞核出现强阳性反应，表明核内的同源ＤＮＡ与地高辛标记的ＲＥＤ－２０ｃＤＮＡ探针杂交。但在睾丸和肝脏内未见杂交信号，提示ＢＥ－２０是附睾尾和近端输精管上皮细胞合成的蛋白质。经分析证明，此蛋白是一种细胞外的蛋白酶抑制剂，可能与防止精子过早获能有关，它是否能用于制备抗生育疫苗有待深入的研究。
Rabbit epididymal tail fluid purified by HPLC, the molecular weight of 20kD epididymis-specific protein, named BE-20. BE-20 antibodies have the ability to significantly inhibit human sperm penetration into zona pellucida hamster eggs. A 0.5 kb cDNA fragment encoding BE-20 has been obtained and is named RED-20. The fragment was labeled with digoxin and in situ hybridization with rabbit testis, epididymis, vas deferens and liver tissue to observe the localization and expression of BE-20 in various tissues. In situ hybridization was performed using both thermal denaturation before and without thermal denaturation. Hybridization signals were purple-brown particles, distributed in the cytoplasm of the main cells of the epididymal tail and epithelial cells of the proximal vas deferens, indicating that the cytoplasmic mRNA hybridized with the digoxigenin labeled RED-20 cDNA probe. Tissue heat-denatured, the epididymal and proximal seminiferous epithelial cell nucleus strong positive reaction, indicating homologous nuclear DNA and digoxin labeled RED-20cDNA probe hybridization. However, no signal was observed in the testis and liver, suggesting that BE-20 is a protein synthesized by the epididymal tail and proximal seminiferous epithelium. The analysis proved that this protein is an extracellular protease inhibitor, may be related to the prevention of premature sperm capacitation, whether it can be used to prepare anti-fertility vaccine needs further study.