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Nogo-66多克隆抗体的制备和鉴定 被引量：1

Preparation and identification of Nogo-66 polyclonal antibodies
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摘要 目的制备抗中枢神经系统(CNS)突起再生抑制因子Nogo胞外段Nogo-66的多克隆抗体,以进行Nogo分子检测和功能研究。方法采用原核系统表达来源于大鼠的GST-Nogo-66融合蛋白,经纯化后免疫新西兰兔制备多克隆抗体,采用免疫印迹、免疫组织化学和细胞培养技术对该抗体的特异性及生物学活性进行鉴定。结果获得了高效价的1∶10000兔抗大鼠Nogo-66多克隆抗体,该抗体能够识别原核表达的Nogo分子,大鼠脊髓切片的免疫组织化学结果显示,Nogo分子在神经元和少突胶质细胞中有广泛表达;并能阻断Nogo分子对PC12细胞突起生长的抑制作用,具有一定的生物学活性。结论制备了能识别天然Nogo分子,并具有良好生物学活性的抗Nogo-66多克隆抗体,为检测Nogo分子和进一步研究其功能提供了有力的工具。 Objective To prepare polyclonal antibodies of anti Nogo-66, the extracellular region of one central nervous system neurite regeneration inhibitor Nogo, which could be used to further identification and functional study of Nogo molecule. Methods Preparing rabbit anti rat Nogo-66 polyclonal antibodies with a purified Nogo-66 fusion protein expressed in E. coli system. Studying its specificity by Western-blot and immuno-histochemical techniques and identifying its biological activity in PC12 cells. Results The high titer （1：10 000） anti rat Nogo-66 polyclonal antibodies were obtained. This antibody could specifically recognize the Nogo protein expressed in E. coli system. Immuno-histochemical staining indicated that the Nogo was widely expressed in rat spinal cord neurons and oligodendrocytes. It could effectively block the neurite extensioninhibition of Nogo protein in PC12. Conclusion Successful preparation of anti rat Nogo polyclonal antibodies provides a useful tool in identification or further functional study of Nogo molecule.
作者 谢云峥 于盼盼 刘东银 陆佩华 黄立东
机构地区 第二医科大学神经生物学实验室
出处 《中华检验医学杂志》 CAS CSCD 北大核心 2005年第10期1064-1067,共4页 Chinese Journal of Laboratory Medicine
基金 上海市科技发展基金资助项目(02JC14014)
关键词 神经再生 抗体 多克隆 抗体 特异性 Nogo-66 Neural regeneration Antibody, polyclonal Antibody, specificity Nogo-66
分类号 R392 [医药卫生—免疫学]
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参考文献9

  • 1Chen MS, Huber AB, van der Haar ME, et al. Nogo-A is a myelin-associated neurite outgrowth inhibitor and an antigen for monoclonal antibody IN-I. Nature, 2000, 403: 434-439.
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同被引文献17

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中华检验医学杂志
2005年 第10期

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