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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Light-emitting diode-induced fluorescence detection of native proteins in capillary electrophoresis.
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Light-emitting diode-induced fluorescence detection of native proteins in capillary electrophoresis.

机译:发光二极管诱导的毛细管电泳中天然蛋白的荧光检测。

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摘要

A continuous-wave 280 nm light-emitting diode (LED) was used as the excitation source for native fluorescence detection of proteins in CE. The operating current and temperature of the LED were optimized in order to achieve high luminescence power. It was found that a forward current of 30 mA and a temperature of approximately 5 degrees C gave the best S/N. By using a set of two ball lenses to focus light from the LED, we achieved a spot of approximately 200 mum with a power of 0.1-0.2 mW on the detection window. Fluorescence was collected with a ball lens at 90 degrees angle through a bandpass filter onto a photomultiplier tube. In CZE an LOD of 20 nM for conalbumin was reached. In capillary gel electrophoresis all eight proteins from a commercial standard kit were detected with high S/N. For a 10 mug/mL total protein mixture, S/N was better than 3 for all proteins in solution. Further improvement in LOD should be possible on utilization of an LED with higher luminescence power.
机译:280 nm连续波发光二极管(LED)被用作激发源,用于CE中蛋白质的天然荧光检测。优化了LED的工作电流和温度,以实现高发光功率。发现正向电流为30 mA,温度约为5摄氏度,可获得最佳的信噪比。通过使用一组两个球形透镜聚焦来自LED的光,我们在检测窗口上获得了约200 mum的光斑,功率为0.1-0.2 mW。用球镜以90度角通过带通滤光片将荧光收集到光电倍增管上。在CZE中,伴白蛋白的LOD达到20 nM。在毛细管电泳中,以高信噪比检测到了来自商业标准试剂盒的所有八种蛋白质。对于10杯/毫升的总蛋白质混合物,溶液中所有蛋白质的S / N均优于3。利用具有更高发光功率的LED,可以进一步提高LOD。

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