一种针对Naked2产生的单克隆抗体。
A monoclonal antibody produced against Naked2.
作者信息
Cao Chang, Wang Shengyu, Lv Sha, Li Zhe, Wang Xianjiang, Zeng Fanwei, Zhang Haipeng, Dai Yujuan, Dou Xiaofeng, Chen Xiaoli, Li Xiudong, Luo Lingli, Hu Tianhui, Yan Jianghua
机构信息
Cancer Research Center, Medical College, Xiamen University, Xiamen, Fujian, China.
出版信息
Monoclon Antib Immunodiagn Immunother. 2013 Aug;32(4):290-4. doi: 10.1089/mab.2012.0111.
Naked2 (NKD2) is a member of the Naked family and negatively regulates canonical Wnt signaling. NKD2 may play a role in embryo development and tumor formation by affecting Wnt signaling. In the present study, we describe the establishment of a monoclonal antibody against NKD2 (anti-NKD2 MAb) through the hybridoma method. The purified anti-NKD2 MAb measured a titer of 2.56 × 10(5) against NKD2 by indirect ELISA. Western blot analysis, immunoprecipitation, and confocal microscope showed that the anti-NKD2 MAb can specifically combine NKD2 protein in SW480 and LOVO cells. Competitive inhibition assays of Western blot and indirect ELISA showed that the anti-NKD2 MAb can be blocked with NKD2(1-217) protein. The anti-NKD2 MAb would be helpful for further studies on the structure activity relationship, protein detecting, and cell-signaling pathway of NKD2.
Naked2（NKD2）是Naked家族的一员，对经典Wnt信号通路起负向调控作用。NKD2可能通过影响Wnt信号在胚胎发育和肿瘤形成中发挥作用。在本研究中，我们描述了通过杂交瘤方法制备抗NKD2单克隆抗体（抗NKD2 MAb）的过程。通过间接ELISA法测定，纯化后的抗NKD2 MAb针对NKD2的效价为2.56×10⁵。蛋白质印迹分析、免疫沉淀和共聚焦显微镜检测表明，抗NKD2 MAb可在SW480和LOVO细胞中特异性结合NKD2蛋白。蛋白质印迹和间接ELISA的竞争抑制试验表明，抗NKD2 MAb可被NKD2（1 - 217）蛋白阻断。抗NKD2 MAb将有助于进一步研究NKD2的结构活性关系、蛋白质检测及细胞信号通路。
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