六种抗大鼠Nogo-A分子单克隆抗体的免疫荧光组织化学染色特性的比较  

Identification of six kinds of monoclonal antibodies against rat Nogo-A molecule in the immunofluorescent histochemical staining

作　　者：邓斌[1] 刘芳芳[1] 赵湘辉[1] 于才勇[1] 卞干兰[1] 冯睿[1] 鞠躬[1] 王键[1] 

机构地区：[1]第四军医大学神经生物学教研室,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2011年第4期425-427,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家重大新药创制大平台项目子课题(2009ZX09301-009-PT02);国家自然科学基金资助项目(30800325)

摘　　要：目的:检测和比较6种针对大鼠Nogo-A分子不同表位的单克隆抗体(mAb)应用于中枢神经组织的免疫组化染色特性,从而决定它们今后可能的应用范围和方法。其中4种mAb是针对Nogo-A分子中Nogo66片段,分别命名为No-go66-1、Nogo66-2、Nogo66-3和Nogo66-4;其他2种mAb是针对Nogo-A分子氨基端(570-691)肽段,分别为Nogo-N1和Nogo-N2。方法:利用免疫荧光组织化学,检测Nogo-A mAb抗体对大鼠神经组织的反应性与特异性。结果:正常SD大鼠脊髓组织的免疫荧光组织化学双标结果显示,制备的6种抗大鼠Nogo-A分子mAb均可以与商品化的兔抗大鼠Nogo-A多克隆抗体(PcAb)双标记;其中Nogo66-1、Nogo66-2、No-go66-4和NogoN-2 mAb可与MBP阳性细胞双标记,与GFAP阳性细胞不共存;而Nogo66-3和NogoN-1 mAb不仅可以与MBP阳性细胞双标,同时也可以和GFAP阳性细胞共存。结论:Nogo66-1、Nogo66-2、Nogo66-4和NogoN-2 mAb可很好的识别组织中的Nogo-A分子,用于免疫组化研究。AIM：To detect and characterize of 6 monoclonal antibodies（mAbs） against different epitopes of rat Nogo-A molecule in immunohistochemistry to decide their applications in futrue.Four mAbs against Nogo66 fragment are named Nogo66-1,Nogo66-2,Nogo66-3 and Nogo66-4.The rest of 2 mAbs against N-termial 570-691aa fragment are named NogoN-1 and NogoN-2.METHODS： The immunofluorescence staining was used to detect the reactivity and specificity of those 6 mAbs in spinal tissue sections of rat.RESULTS： All 6 mAbs were double-labelled with commercial rabbit anti-Rat Nogo-A polyclonal antibody（PcAb） in spinal cord sections respecitvely.All 6 mAbs were colocalization with MBP respectively.However Nogo66-3 and NogoN-1 could also be double-staining with GFAP respectively.CONCLUSION： Nogo66-1,Nogo66-2,Nogo66-4 and NogoN-2 could recognize specifically in Nogo-A protein of tissues in immunohistochemical methods.

关 键 词：NOGO-A 单克隆抗体 免疫荧光 组织化学 

分 类 号：Q813.2[生物学—生物工程] R392.11[医药卫生—免疫学]