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超灵敏蛋白质检测与成像:Lumitein™、ProteoSilver™、SYPRO® Ruby和考马斯亮蓝凝胶染色剂的比较

Ultrasensitive protein detection and imaging: comparison of Lumitein™, ProteoSilver™, SYPRO(®) Ruby, and Coomassie (®) Brilliant Blue gel stains.

作者信息

Ren Guang, Okerberg Carl K, Mathews Suresh T

机构信息

Boshell Diabetes and Metabolic Diseases Research Program, Auburn University, Auburn, AL, USA.

出版信息

Methods Mol Biol. 2012;869:621-32. doi: 10.1007/978-1-61779-821-4_57.

Abstract

Following sodium dodecyl sulfate polyacrylamide gel electrophoresis, proteins can be visualized by various methods of detection and imaging. Traditional methods of protein gel detection and imaging have been improved and expanded through technological advancement. Today, the detection of proteins, resolved on gels, can be accomplished with a variety of stains with various sensitivities. Digital cameras used in the imaging of protein gels are not only more sensitive than their film precursors, but they can be used in combination with imaging software that offers a host of useful applications. Here we describe the UVP BioImaging System in combination with LabWorks Image and Acquisition software to provide a comparison of four different protein gel stains: Lumitein™, ProteoSilver™, SYPRO(®) Ruby, and Coomassie(®) Brilliant Blue. We demonstrate that the detection sensitivity limit appears to be between 100 and 500 ng/protein band of protein with Coomassie(®) Brilliant Blue, 10-50 ng/protein band with Lumitein™ and SYPRO(®) Ruby, and as little as 5 ng/protein band with the ProteoSilver™ stain.

摘要

在十二烷基硫酸钠聚丙烯酰胺凝胶电泳之后,蛋白质可以通过各种检测和成像方法进行可视化。传统的蛋白质凝胶检测和成像方法通过技术进步得到了改进和扩展。如今,对凝胶上分离出的蛋白质进行检测可以使用多种具有不同灵敏度的染色剂来完成。用于蛋白质凝胶成像的数码相机不仅比其胶片前身更灵敏,而且可以与提供大量有用应用程序的成像软件结合使用。在这里,我们描述了UVP生物成像系统与LabWorks图像采集软件相结合,以比较四种不同的蛋白质凝胶染色剂:Lumitein™、ProteoSilver™、SYPRO® Ruby和考马斯亮蓝。我们证明,考马斯亮蓝对蛋白质的检测灵敏度极限似乎在每条蛋白带100至500纳克之间,Lumitein™和SYPRO® Ruby为每条蛋白带10至50纳克,而ProteoSilver™染色剂低至每条蛋白带5纳克。

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