High-throughput proteomics processing of proteins in polyacrylamide in a multiwell format
High-throughput proteomics processing of proteins in polyacrylamide in a multiwell format
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DOI：
10.1021/pr060472y
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发表时间：
2007-01-01
影响因子：
4.4
通讯作者：
Hilliard, George M.
中科院分区：
文献类型：
--
作者：
Cummings, Edwin D.;Brown, Janet M.;Hilliard, George M.
在高灵敏度下从聚丙烯酰胺中处理多个蛋白质样品是提高大容量蛋白质鉴定项目成功率的主要瓶颈。使用蛋白质标准品对用于处理聚丙烯酰胺中的蛋白质的多孔滤板方法的灵敏度进行了优化。结果表明，该方法在凝胶加载范围内是一种可靠且可重现的方法，适用于鉴别接近银染色阈值的蛋白质。这种高通量手动方法需要最少的专用设备，并且可以与蛋白质组学基础设施断开连接，用于制备质谱准备样品。
Processing multiple protein samples from polyacrylamide at significant sensitivity represents a major chokepoint for raising the success rate in high-volume protein identification projects. A multiwell filterplate method for processing proteins in polyacrylamide was optimized for sensitivity using a protein standard. The results demonstrate this process to be a reliable and reproducible method over a range of gel loadings and suitable for the identification of proteins near the threshold of silver stain. This high-throughput manual method requires a minimum of specialized equipment, and can be performed disconnected from a proteomics infrastructure for the preparation of mass spectrometry-ready samples.