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Identification of proteins from human cerebrospinal fluid, separated by two-dimensional polyacrylamide gel electrophoresis

机译:二维聚丙烯酰胺凝胶电泳分离鉴定人脑脊髓液中的蛋白质

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The aim of this work is to display the protein composition of the cerebrospinal fluid by two-dimensional (2-D) gel electrophoresis and identify it using different mass spectrometric techniques. This will enable us to present an overview of the proteins in human cerebrospinal fluid. The comparison of 2-D gels will help us to analyze the normal protein variability in healthy persons and specific protein variations in patients with different neurological diseases (e.g., morbus Alzheimer, chorea Huntington). However, it is not possible to carry out 2-D gel electrophoresis directly with human cerebrospinal fluid due to the high amount of salts, sugars and lipids present. In addition, the total amount of protein is only as high as 0.3-0.7 mu g/mu L. Therefore, concentration and desalting steps using precipitation and ultrafiltration are necessary. To date we have been able to identify more than 65 spots from 2-D gels using matrix assisted laser desorption/ionization-mass spectrometry and electrospray ionization-mass spectrometry. [References: 19]
机译:这项工作的目的是通过二维(2-D)凝胶电泳显示脑脊髓液的蛋白质组成,并使用不同的质谱技术对其进行鉴定。这将使我们能够概述人脑脊髓液中的蛋白质。二维凝胶的比较将有助于我们分析健康人的正常蛋白质变异性以及患有不同神经系统疾病(例如morbus Alzheimer,chorea Huntington)的患者的特定蛋白质变异。但是,由于存在大量的盐,糖和脂质,因此不可能直接用人脑脊髓液进行2-D凝胶电泳。另外,蛋白质的总量仅高达0.3-0.7μg/μL。因此,需要使用沉淀和超滤的浓缩和脱盐步骤。迄今为止,我们已经能够使用基质辅助激光解吸/电离质谱和电喷雾电离质谱从二维凝胶中鉴定出65个以上的斑点。 [参考:19]

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