Control of the Fluorescence of Dye-Antibody Conjugates by (2-Hydroxypropyl)-β-cyclodextrin in Fluorescence Microscopy and Flow Cytometry
Control of the Fluorescence of Dye-Antibody Conjugates by (2-Hydroxypropyl)-β-cyclodextrin in Fluorescence Microscopy and Flow Cytometry
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DOI：
10.1021/ac2014146
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发表时间：
2011-09-15
影响因子：
7.4
通讯作者：
Dubrovsky, Tim
中科院分区：
文献类型：
--
作者：
Guryev, Oleg;Abrams, Barny;Dubrovsky, Tim
当蛋白质与荧光有机染料缀合时，染料分子的荧光发射通常降低，有时高达50- 70%。这种猝灭现象已经被认识了几十年，但是到目前为止，还没有简单实用的方法来控制与蛋白质缀合的染料的荧光，特别是对于与免疫球蛋白缀合的染料。在这里，我们报告说，除了（2-羟丙基）-β-环糊精（HP β CD）的染料抗体结合物可以增加荧光高达2.5倍，在细胞成像和流动分析。这种方法可能是一种有效的方法，以提高检测灵敏度的荧光有机标记用于免疫学，组织化学和细胞生物学。
When proteins are conjugated to fluorescent organic dyes, fluorescence emission of the dye molecules is usually decreased, sometimes up to 50-70%. This quenching phenomenon has been acknowledged for decades, but as yet, there are no simple, practical methods to control the fluorescence of dyes conjugated to proteins, especially for dyes conjugated to immunoglobulins. Here, we report that the addition of (2-hydroxypropyl)-beta-cyclodextrin (HP beta CD) to dye-antibody conjugates can increase fluorescence up to 2.5-fold in cell imaging and flow analysis. This method may be an effective way to increase the sensitivity of detection of fluorescent organic labels used in immunology, histochemistry, and cell biology.