Preparation of Polyclonal Antibodies Against Testis-specific Protease 50 and Characterization of Antibody Specificity

摘要

Testis-specific protease 50 (TSP50) is a testis-specific oncogene,which is abnormally activated in most tested patients with breast cancer.This property makes it an attractive molecular marker and a promising target for the diagnosis and therapy of breast cancer.In order to obtain the protective and specific polyclonal antibodies for further research ,TSP50 cDNA was amplified by RT-PCR from normal human testicular tissue,and inserted into eukaryotic expression vector pcDNA3.1.Rabbit anti-TSP50 polyclonal antibodies were prepared by means of intramuscular injection of pcDNA3.1-TSP50 into the rabbits.Titers of the anti-sera were measured by ELISA and Western blotting with the E.coli cell lysate containing the induced CST-TSP50 fusion protein as an antigen.In addition,we examined the expression of TSP50 in both breast cancer cell line MCF-7 and breast cancer tissue by immunofluorescent and immu-nohistochemistry analysis.