胶质纤维酸性蛋白和神经微丝蛋白的纯化及其抗体的制备和鉴定

SIMULTANEOUS PURIFICATION OF GLIAL FIBRILLARY ACIDIC PROTEIN AND NEUROFILAMENT,PREPARATION AND IDENTIFICATION OF THE TWO CORRESPONDING ANTIBODIES

参考Ｅｎｇ和Ｌｉｅｍ两法予以改进。自牛脊髓中同时提取纯化胶质纤维酸性蛋白和神经微丝蛋白，用淋巴结内微量注射法免疫家兔获得相应抗体。用免疫转移电泳和免疫组织化学技术（ＰＡＰ法）对抗体进行鉴定。兔抗胶质纤维酸性蛋白与牛脊髓的相应蛋白组分以及中枢神经系统和视神经的星形胶质细胞呈阳性反应。兔抗神经微丝蛋白则和牛脊髓中的相应蛋白以及神经元和神经纤维呈阳性反应。

Glial fibrillary acidic protein(GFAP) and neurofilament(NF) were purified from bovine spinal cord according to the methods from Eng and Liem with modification. Anti-GFAP and Anti-NF were raised in rabbits by injection of micrograms of the antigens into lymph nodes. The antibodies were identified by immunoblot and immunohistochemical staining(PAP method). The results showed that anti-GFAP reacted with GFAP band on nitrocellulose sheet,astrocytes of central nervous system and optical nerve, while anti-NF reacted with neurofilament bands on nitrocellulose sheet, neurons, neurites and dendrites.