当前位置: 首页 > 文章 > Immune Blot Analysis on Expression of the Mammalian Target of Rapamycin in Goat Fetal Fibroblasts with Recombinant Polyclonal Antibody 农业科学学报 (英文) 2012,11 (6)
Position: Home > Articles > Immune Blot Analysis on Expression of the Mammalian Target of Rapamycin in Goat Fetal Fibroblasts with Recombinant Polyclonal Antibody Journal of Integrative Agriculture 2012,11 (6)

Immune Blot Analysis on Expression of the Mammalian Target of Rapamycin in Goat Fetal Fibroblasts with Recombinant Polyclonal Antibody

作  者:
Liang Yan;Li Shu-yu;Wang Xiao-jing;Wu Man-lin;Yang Jiao-fu;Li Jie;Hao Xi-yan;Hao Hui-fang;Ba Yin-ma;Chen Xian-wei;Wang Zhi-gang
单  位:
Inner Mongolia Univ, Coll Life Sci, Hohhot 010021, Peoples R China
关键词:
prokaryotic expression;recombinant antibody;goat;mTOR;CCI-779
摘  要:
To detect the mammalian target of rapamycin (mTOR) expressed in Cashmere goat fetal fibroblasts (GFb), mTOR gene was cloned from Inner Mongolia Cashmere goat (Capra hircus) and expressed in Escherichia coli followed by immunizing mice with the purified recombinant protein as an immunogen to produce the anti-goat mTOR recombinant polyclonal antibody. Antiserum was collected from the immunized mice after the fifth immunization and its titer was determined with enzyme-linked immunosorbent assay (ELISA). The results showed that the recombinant polyclonal antibody had a titer 1:200000 and could react with the mTOR expressed in GFb cells with a specific and sensitive affinity. Western blot showed that mTOR expression and phospho-mTOR (Ser 2448) activity were inhibited when GFb cells were treated with CCI-779, an mTOR specific inhibitor.

相似文章

计量
文章访问数: 11
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊