Expression, purification of the recombinant protein fragment of the monkey epididymis expressed gene ESc-615 and preparation of polyclonal antisera to it

Document Type

Article

Publication Date

12-1-2002

Abstract

ESc-615 is a gene which was specifically expressed in epididymis, newly discovered in our laboratory. To elucidate the function of ESc-615 in sperm maturation and fertilization at the protein level, its C-terminal 310 amino acid residues fragment was expressed in E. coli and used to immunize rabbits to obtain polyclonal antiserum with titer of 100 000. The antisera could detect the antigen as low as 3 ng. A 63 kD protein in rat epididymis protein extracts could be detected in Western blot by using this particular antiserum, in accordance with the size of ESc-615 homologue polypeptide deduced from its cDNA. Immunostaining results showed that ESc-615 was a secreted protein and could bind sperms, which was similar with other carboxylesterases in male reproductive system; the positive staining was eliminated after antigen absorption. All these results showed that this antiserum was not only effective but also highly specific, providing a useful tool in further functional studies of ESc-615.

Publication Source (Journal or Book title)

Acta Biochimica et Biophysica Sinica

First Page

753

Last Page

756

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