摘要
本文采用羟基磷灰石吸附洗脱法,从牛脊髓中分离神经微丝(NF)。用SDS-PAGE显示其分子量为72K,164K及195K,负染电镜可见直径为8-10nm的丝状结构。以此为抗原免疫BALB/c小鼠,取其脾细胞与NS1细胞融合,获得了一株持续分泌抗NF单克隆抗体的杂交瘤细胞株AN_1。其染色体众数为91,所分泌抗体属IgG1亚类。免疫印迹表明此抗体为NF195特异性。免疫组化显示该抗体具有神经纤维特异性,对神经原胞体及非神经组织呈阴性反应。
Hydroxyapatite adsorption chromatography was used to extract neurofila-ments from bovine spinal cords.SDS-PAGE showed that it had molecular weights of 72K, 164K, 195K. Negative stainiUg electron microscopy revealed it is an intermediate filament of 8-10 nm in diameter. A hybridoma cell line AN1 continuously secreting anti-neurofilament monoclonal antibody was established by the fusion of mouse myeloma cell line NSl and spleen cell from BALB/c mice immunized with neurofilament extracts. Karyotype analysis shoved that the chromosome mode was 91. The immunodif fusion test showed that ANl McAb belongs to IgGl subclass. The immunoblotting test indicated that the McAb is specific for NF195. The immunohistochemical examination showed that the McAb has a specificity for the axon fibers, but the neurons of normal and neuplastic neural tissues, non-neural tissues and tumors are negative. It can produce high titer McAb after intraperitoneal injection in BALB/c mice, and can secrete the McAb after freezing in liquid nitroeen and thawing.
出处
《免疫学杂志》
CAS
CSCD
北大核心
1990年第2期92-96,共5页
Immunological Journal
关键词
神经微丝
单克隆抗体
制备
鉴定
Neurofilament, Hybridoma technique, Monoclonal antibody,Nerve fibers
