多疣壁虎SPARCL1多克隆抗血清的制备及鉴定  

Preparation and identification of polyclonal antibody against Gekko japonicus SPARCL1

作　　者：周晓芳[1] 陈倩[1] 储钰丹[1] 王学谦[1] 钱天梅[1] 陆仁飞[1] 顾星星[1] 

机构地区：[1]南通大学神经再生重点实验室,南通226001

出　　处：《南通大学学报（医学版）》2011年第2期79-82,86,共5页Journal of Nantong University(Medical sciences)

基　　金：国家自然科学基金资助项目(30671046;31071288)

摘　　要：目的:制备针对多疣壁虎SPARCL1的多克隆抗体,为深入研究SPARCL1基因功能奠定基础。方法:构建pGEX-4T1-SPARCL1质粒,在大肠杆菌BL21表达GST-SPARCL1蛋白,经亲和层析纯化后免疫新西兰大白兔,获得SPARCL1抗血清,经酶联免疫吸附试验(ELISA)法检测SPARCL1抗血清的效价,Western blot和免疫组化法检测多克隆抗体特异性。结果:成功构建多疣壁虎SPARCL1基因的原核表达质粒pGEX-4T1-SPARCL1,诱导得到较高纯度的GST-SPARCL1融合蛋白,免疫兔获得SPARCL1抗血清,经Western-blot及免疫组织化学实验显示所得抗体具有较高的效价及特异性。结论:成功获得较高效价特异性强的SPARCL1抗血清,为进一步深入研究多疣壁虎SPARCL1功能提供抗体工具。Objective： To prepare the polyclonal antibody against Gekko japonicus SPARCL1 for future functional studies of SPARCL1.Methods： Prokaryotic expression vector pGEX-4T1-SPARCL1 was transformed into E.coli BL21.The recombination proteins were purified using Glutathione Sepharose.The purified fusion proteins were inoculated into adult rabbits to develop antiserum.Enzyme-linked ELISA,Western blot and immunohistochemistry staining were used to evaluate the specificity of the prepared antiserum.Results： Prokaryotic expression vector pGEX-4T1-SPARCL1 was successfully expressed.The fusion protein was used to immunize rabbits.Immunohistochemical and Western blot showed a high specificity of the antiserum of g-SPARCL1.Conclusion： We successfully amplified and expressed the g-SPARCL1 in E.coli.The obtained antiserum of g-SPARCL1 showed a high specificity.The proteins and antiserum prepared in this study can be used for further research of the function investigation of Gekko SPARCL1.

关 键 词：SPARCL1 融合蛋白 多疣壁虎 抗血清 

分 类 号：R392.11[医药卫生—免疫学]