NBD-SCN 对半胱氨酸/同型半胱氨酸的灵敏度评价及其生物成像应用。
Sensitivity evaluation of NBD-SCN towards cysteine/homocysteine and its bioimaging applications.
机构信息
Department of Biological Science and Technology, Institute of Molecular Medicine and Bioengineering, National Chiao Tung University, 75 Bo-Ai Street, Hsinchu 300, Taiwan.
Department of Biological Science and Technology, Chung Hwa University of Medical Technology, 89, Wunhua 1st Street, Rende, Tainan 717, Taiwan.
出版信息
Biosens Bioelectron. 2014 Jun 15;56:117-23. doi: 10.1016/j.bios.2014.01.009. Epub 2014 Jan 11.
A push-pull fluorogenic reagent, NBD-SCN, was applied for specific detection of cysteine (Cys) and homocysteine (Hcy). Replacing thiocyanato group with Cys/Hcy increased the push-pull characteristic of the probe and resulted in emission of fluorescence. The fluorescent response of the probe toward Cys/Hcy was significantly higher than toward glutathione and other amino acids. The probe showed a 470- and 745-fold fluorescence enhancement at 550 nm and detection limit of 2.99 and 1.43 nM for Cys and Hcy, respectively. Time-dependent fluorescence assays showed that the fluorescence intensity reached a plateau within 20s after addition of Cys and within 10 min after addition of Hcy. Furthermore, the fluorescence images of Cys/Hcy in Raw 264.7 cells were obtained after adding this probe to the cells. These results indicate that NBD-SCN not only possesses good selectivity and sensitivity for Cys/Hcy but also can penetrate cells for Cys/Hcy bioimaging.
一种推拉式荧光试剂 NBD-SCN 被应用于半胱氨酸 (Cys) 和高半胱氨酸 (Hcy) 的特异性检测。用 Cys/Hcy 取代硫氰酸根，增加了探针的推拉特性，导致荧光发射。探针对 Cys/Hcy 的荧光响应明显高于对谷胱甘肽和其他氨基酸的响应。该探针对 Cys/Hcy 的荧光增强倍数分别为 470 倍和 745 倍，检测限分别为 2.99 和 1.43 nM。时间依赖性荧光分析表明，加入 Cys 后 20s 内荧光强度达到平台，加入 Hcy 后 10min 内荧光强度达到平台。此外，将该探针加入 Raw 264.7 细胞后，可获得细胞内 Cys/Hcy 的荧光图像。这些结果表明，NBD-SCN 不仅对 Cys/Hcy 具有良好的选择性和灵敏度，而且可以穿透细胞进行 Cys/Hcy 生物成像。
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