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Company: Thorlabs
Catalog#: PT3
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Simultaneous Imaging of Single Protein Size, Charge, and Binding Using A Protein Oscillation Approach
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2021-03-05
[Abstract]  

Electrophoresis and Western blot are important tools in protein research for detection and identification of proteins. These traditional techniques separate the proteins based on size and charge differences and identify the proteins by antibody binding. Over the past decade, the emergence of single-molecule techniques has shown great potential in improving the resolution of the traditional protein analysis methods to the single-molecule level. However, such single-molecule techniques measure either size or charge, and it is challenging to measure both at the same time. Recently, we have developed a single-molecule approach to address this problem. We tether the single proteins to a surface with a polymer linker and drive them into oscillation with an electric field. By tracking the

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[摘要]  [摘要]电泳和蛋白质印迹是蛋白质研究中用于检测和鉴定蛋白质的重要工具。这些传统技术根据大小和电荷差异分离蛋白质,并通过抗体结合鉴定蛋白质。Ø版本在过去的十年中,单分子技术的出现显示了将传统蛋白质分析方法的分辨率提高到单分子水平的巨大潜力。然而,这样的单分子技术测量大小或电荷,并且同时测量这两者是挑战性的。最近,我们已经开发出一种单分子方法来解决这个问题。我们使用聚合物接头将单个蛋白质束缚在表面上,并通过电场驱动它们振荡。通过使用光学成像方法跟踪蛋白质对电场的机电响应,可以同时获得大小和电荷。抗体或离子与拴系蛋白的结合也会改变大小和电荷,这使我们能够探查相互作用。该协议包括蛋白质振荡器的制造,光学检测系统的配置以及用于定量蛋白质大小和电荷的振荡信号的分析。我们希望该协议将使研究人员能够在单个平台上执行全面的单蛋白分析。



[背景]蛋白质在许多生物学过程的发挥重要作用,并作为药物靶标和生物标志物。蛋白质分析依赖于电泳,蛋白质印迹和质谱等技术,这些技术根据蛋白质分子的大小和电荷来分离和鉴定蛋白质。尽管这些技术在检测蛋白质方面功能强大,但它们对阐明分子异质性和精确诊断所必需的单个分子不敏感。最近,已经开发了几种单分子技术来测量单个生物分子的大小或电荷。实例包括:i nterferometric散射显微镜(iSCAT ...

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