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.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
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)
