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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 To prepare a monoclonal antibody against 43k D protein of peripheral nerve and detect its expression in normal and injured sciatic nerve. Methods The SDS-polyacrylamide gel electrophoresis system was used to separate and recover the 43kD protein from the peripheral nerve. The BALB / c mice were immunized by hybridoma technology and dot blot assay to obtain 43kD protein The specificity of the monoclonal antibody was detected by Western blot and the expression of 43kD protein in the normal sciatic nerve and injured distal sciatic nerve was detected. Results A monoclonal antibody against 43kD protein was detected. Western blot showed that a specific positive reaction band appeared at 43kD in the distal sciatic nerve distal sciatic nerve of normal rats and sciatic nerve 2 weeks after injury. In the injured nerve The 43k D protein positive reaction product is colored darker. Conclusion 43k D protein has unique immunochemical characteristics, which are expressed both in normal and injured sciatic nerves, and stronger in injured sciatic nerve.