利用压力循环技术(PCT)对皮质骨中分离出来的蛋白和多肽进行电泳分析

2008/07/31   下载量: 84

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应用领域
检测样本
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Extraction of proteins from extensively calcified osseous tissue, such as cortical bone has been particularly challenging for traditional methods of sample preparation. However, a comprehensive proteomic analysis of bone is only possible when the total protein constituency is effectively isolated. The efficiency of sample preparation is therefore a critical component of the analytical process. Historically, extraction of protein from bone required prolonged acid demineralization over several days to enable complete penetration of histochemical reagents to cellular components. Here we describe a method for the extraction of protein from ostrich tibia, which was used as a model sample to develop an extraction process that uses pressure cycling technology (PCT) and also which obviates the need for acid demineralization prior to extraction. The ability to extract proteins from bone without prior demineralization offers important advantages in efficient representative extraction of protein and significant time savings during sample preparation. 利用压力循环技术(PCT)对皮质骨中分离出来的蛋白和多肽进行电泳分析 从皮质骨等充分钙化的骨质组织中提取蛋白的方法极大地挑战了传统的样品制备方法。然而这种方法只有在总的目标蛋白有效分离的时候,广泛的蛋白质组学分析才能成为可能。因此,样品制备的有效性成为分析过程中的关键因素。历史上,从骨头中提取蛋白需要几天的去除矿物质的过程使得组织化学试剂完全渗透到细胞组分中。本文描述了一种从鸵鸟胫骨中提取蛋白的方法,该方法利用了压力循环技术(PCT),而且省去了以往的在提取蛋白之前先对样品进行去除矿物质的过程。这种方法使得提取蛋白非常有效而且节约了大量的时间成本。

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