摘要
目的 制备周围神经 43k D蛋白单克隆抗体 ,并检测该蛋白在正常及损伤坐骨神经中的表达。方法 实验用SDS-聚丙烯酰胺凝胶电泳系统 ,从周围神经中分离回收 43k D蛋白作为抗原 ,免疫 BAL B/ c小鼠 ,通过杂交瘤技术和点膜印迹法检测 ,获得分泌识别 43k D蛋白的单克隆抗体的杂交瘤细胞株 ,以 Western blot方法检测单克隆抗体的特异性 ,并检测43k D蛋白在正常坐骨神经及损伤坐骨神经远侧端中的表达。结果 经检测获得了识别 43k D蛋白的单克隆抗体 ,Westernblot显示在正常大鼠坐骨神经与损伤后 2周的坐骨神经远侧端组织电泳图谱 43k D处均出现特异的阳性反应条带 ,在损伤神经中 43k D蛋白阳性反应产物着色较深。结论 43k D蛋白具有独特的免疫化学特性 ,在正常与损伤坐骨神经中均有表达 ,在损伤坐骨神经中表达更强。
? Objective The monoclonal antibody against 43kD protein of peripheral nerves was prepared and the expression of 43kD protein in injured sciatic nerves was evaluated. Method The 43kD protein, as an antigen, was isolated from peripheral nerves by SDS PAGE. The monoclonal antibody was prepared by immunizing the BALB/c mouse and by dot blot assay. Western blotting was employed to demonstrate the specificity of the monoclonal antibody and to exam the expression of 43kD protein in the distal stump of sciatic nerves transected 2 weeks earlier. Result A special positive band was found on 43kD of normal peripheral nerves and a deep staining positive band on the distal stump of injured sciatic nerves. Conclusion The result demonstrates that the specific 43kD protein is expressed in both normal and injured peripheral nerves; and it is unique in immunochemistry. The expression of the 43kD protein is up regulated in the distal stump of injured sciatic nerves compared with that in normal nerves.\;〔
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2002年第1期1-3,共3页
Chinese Journal of Histochemistry and Cytochemistry
基金
国家自然科学基金资助(No.30070255)
