Pixley S K, Kobayashi Y, de Vellis J
J Neurosci Res. 1984;12(4):525-41. doi: 10.1002/jnr.490120402.
A monoclonal antibody was developed using rat astrocytes purified in vitro as the starting antigenic material. Selection of the monoclonal was on the basis of astrocyte binding specificity in brain sections using indirect immunofluorescence techniques. The antibody (RBA2) that was chosen was specific for astrocytes in that it did not stain neurons or oligodendrocytes in frozen brain sections. It did, however, show binding to vascular smooth muscle and meningeal cells. The antigenic determinant(s) was determined to be on filaments of the intermediate-size class in cultured astrocytes and fibroblasts. From analysis of binding patterns in various tissues and in immunoblots, it was found that RBA2 cross-reacted strongly with glial fibrillary acidic protein (GFAP) and desmin. There was a weaker cross-reactivity to a vimentin-associated component. It is proposed that this antibody can be used as an astrocyte and blood vessel marker in brain sections, a vimentin marker in cultures and as a probe of intermediate filament composition and distribution.
以体外纯化的大鼠星形胶质细胞作为起始抗原材料,制备了一种单克隆抗体。该单克隆抗体的筛选基于使用间接免疫荧光技术在脑切片中对星形胶质细胞的结合特异性。所选择的抗体(RBA2)对星形胶质细胞具有特异性,因为它在冷冻脑切片中不染色神经元或少突胶质细胞。然而,它确实显示出与血管平滑肌和脑膜细胞的结合。抗原决定簇被确定存在于培养的星形胶质细胞和成纤维细胞中的中等大小类别的细丝上。通过对各种组织中的结合模式和免疫印迹的分析,发现RBA2与胶质纤维酸性蛋白(GFAP)和结蛋白强烈交叉反应。与波形蛋白相关成分的交叉反应较弱。有人提出,这种抗体可作为脑切片中的星形胶质细胞和血管标记物、培养物中的波形蛋白标记物以及中间丝组成和分布的探针。