Fluorescence polarisation of green fluorescent protein (GFP). A strategy for improved wavelength discrimination for GFP determinations

A. Knight, N. Goddard, P. Fielden, M. Barker, N. Billinton, R. Walmsley
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引用次数: 14

Abstract

The fluorescence of green fluorescent protein (GFP), present both within whole yeast cells and in protein extracts from yeast cells, has been observed to be significantly polarised. Fluorescence polarisation is proposed as a useful technique to allow some discrimination between GFP fluorescence and that of other interfering species in cell or media matrices, which have fluorescence bands that overlap those of GFP, which should lead to improved resolution and limits of detection. The method has been tested by discriminating between the fluorescence of GFP in cell extracts and added fluorescein, both of which fluoresce brightly at the same wavelength. The flow-through instrumentation incorporating an argon-ion laser developed for this work is also described.
绿色荧光蛋白(GFP)的荧光偏振。一种改进GFP测定波长辨别的策略
绿色荧光蛋白(GFP)的荧光,存在于整个酵母细胞和酵母细胞的蛋白质提取物中,已经观察到明显的极化。荧光偏振被认为是一种有用的技术,可以区分GFP荧光和细胞或介质基质中其他干扰物质的荧光带,这些干扰物质的荧光带与GFP的荧光带重叠,这将提高分辨率和检测范围。通过对细胞提取物中GFP的荧光和添加荧光素的荧光进行区分,验证了该方法在同一波长上发出明亮的荧光。本文还介绍了为这项工作开发的带有氩离子激光器的流动仪器。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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