Preparation and characterization of polyclonal antibodies against syncytin
SUN Yi
PANG Wei
OUYANG Dongyun
ZHENG Yongtang
Abstract：Syncytin plays a crucial role in syneytiotrophoblast formation. This study is aimed to generate and identify the rabbit polyclonal antibodies against human syncytin. We amplified the syncytin gene by PCR, and then inserted it into cloning vector. After digestion by BamH Ⅰ/Not Ⅰ, the recomhinants were cloned into expression vector pET30a(+) and transferred into E. coli BL21 (DE3);fusion protein His/syncytin was induced by IPTG, and then analyzed by SDS-PAGE; the protein straps about 42 000 were cut, grind-ed, and then used to immunize New Zealand rabbits. Sera of the immunized rabbits were collected 10 days after the second enhanced immunization; the antiserum was detected by ELISA, Western blot, and immunohistochemistry staining. In this study, the prokaryotic expression vector pET30a (+)/syncytin was successfully constructed and fusion protein Syncytin-His was expressed efficiently. The lev-els of polyclonal antibodies were raised in the immunized rabbits. The syncytin polyclonal antibodies reveal high titer and specificity,which may be applied in syncytin function research.
Keywords：SyncytinProkaryotic expressionPolyclonal antibody
Publication Date：2010-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 124-127,131 )