Fukuyama R, Fushiki S, Fujita S
Department of Pathology, Kyoto Prefectural University of Medicine, Japan.
J Neurosci Methods. 1991 Dec;40(2-3):133-7. doi: 10.1016/0165-0270(91)90062-5.
Glial fibrillary acidic protein (GFAP) was purified from normal bovine brain by a modification of the procedure used to isolate vimentin in order to avoid contamination by other cytoskeletal components: vimentin, neurofilament triplet proteins, tubulin and actin. GFAP is thought to be separated from vimentin in the DE cellulose column chromatography step. The three other major proteins were also separable through ion exchange and gel filtration column chromatographies. A purified 49 kDa polypeptide was estimated to be GFAP from peptide mapping and subsequent immunoblotting analysis. We obtained 4.4 mg GFAP/1 g bovine brain white matter in less than 3 days. The polyclonal antibody raised against purified GFAP was able to detect 49 kDa GFAP by immunoblotting analysis. This isolation method is simpler and more rapid than previous methods.
通过对用于分离波形蛋白的程序进行改进，从正常牛脑中纯化出胶质纤维酸性蛋白（GFAP），以避免受到其他细胞骨架成分的污染，这些成分包括波形蛋白、神经丝三联体蛋白、微管蛋白和肌动蛋白。GFAP被认为是在DE纤维素柱层析步骤中与波形蛋白分离的。其他三种主要蛋白质也可通过离子交换和凝胶过滤柱层析分离。通过肽图分析和随后的免疫印迹分析，估计一种纯化的49 kDa多肽为GFAP。在不到3天的时间里，我们从1 g牛脑白质中获得了4.4 mg GFAP。针对纯化的GFAP产生的多克隆抗体能够通过免疫印迹分析检测到49 kDa的GFAP。这种分离方法比以前的方法更简单、更快速。
