sLRIG1蛋白多克隆抗体的制备及鉴定.pdf

中国临床神经外科杂志2012 年12 月第17卷第12期 Chin J Clin Neurosurg ，December 2012 ，Vol. 17 ，No.12 - 743 - ● 实验研究实验研究● sLRIG1 蛋白多克隆抗体的制备及鉴定 易 伟 余信远 柯应兵 卢祖能 【摘要】目的制备专门针对富含亮氨酸重复区免疫球蛋白样蛋白1（LRIG1 ）胞外段（或称可溶性LRIG1 ，sLRIG1）抗体。方 法人工设计、合成sLRIG1 抗原多肽，免疫日本大耳朵白兔，颈动脉取血，分离兔血清；酶联免疫吸附分析法测定抗体效价，免疫 印迹鉴定所获抗体。结果抗sLRTG1 抗体滴度可高达1∶512 000 ；免疫印迹证实所获抗体能与sLRIG1 特异性结合，在100 kD 处 蛋白显影清晰。结论成功获得兔抗人sLRIG1 多克隆抗体，可满足进一步研究的需要。 【关键词】富含亮氨酸重复区免疫球蛋白样蛋白1（LRIG1 ）；可溶性LRIG1 ；多克隆抗体；抗体制备；鉴定 【文章编号】1009-153X（2012 ）12-0743-03 【文献标志码】A 【中国图书资料分类号】R 456 Preparationandidentificationofthepolyclonalantibodytosolubleleucine-richrepeatsandimmunoglobulin-likedomains1 YIWei*,YUXin-yuan*,KEYing-bing,LUZu-neng.*DepartmentofNeurosurgery,RenminHospital,WuhanUniversity,Wuhan 430060,China 【Abstract】ObjectiveTo obtain the good quality antibody to the soluble leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1). Methods The ectodomains, i.e. soluble LRIG1 (gene No. AF381545), which were designed and synthesized in vitro, were intracutaneously injected in Japanese white rabbits. The antibodies which were obtained from the immunized rabbits ’serum were purified. The obtained antibodies were identified by SDSand western blotting. The titre of the antibodies was measured by enzyme-linked immunoassay. Results The antibodies which could specifically bind to the soluble LRIG1 were successfully obtained. There was a clear protein blot around 100 kD on the western film as expected. The titre of the antibodies was high as 1 ∶512 000. Conclusion The polyclonal antibodies to soluble LRIG1 which were successfully obtained may satisfy the needs of the further experimental research. 【Keywords】Soluble leucine-rich repeats and immunoglobulin-like domains 1; Polyclonal antibody; Preparation; Identifica