Nanjing Maternity and Child Health Care Institute, Women's Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, Nanjing, 210004, People's Republic of China.
Department of Laboratory Medicine, The Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, 210003, People's Republic of China.
Mikrochim Acta. 2019 Dec 5;187(1):22. doi: 10.1007/s00604-019-4025-1.
A method is described for the determination of the CCAAT/enhancer binding protein alpha (C/EBPα) which is a regulator in adipocyte differentiation. The method is based on quenching of the red fluorescence (with excitation/emission maxima at 548/562 nm) of Cy3-labeled DNA if it becomes adsorbed on positively charged gold nanoparticles (AuNPs). Fluorescently labeled dsDNA that can bind C/EBPα is introduced as a fluorescent probes. The dsDNA is electrostatically adsorbed on the positively charged AuNPs to quench their fluorescence. In the presence of C/EBPα, it will bind dsDNA which then diffuses away. The fluorescence of the AuNPs becomes restored. The fluorescent signal increases linearly in the 0.05 to 600 ng·mL μM C/EBPα concentration range, and the detection limit is 29 pg·mL. The method is specific and was applied to analyze cell lysates and in-situ. Graphical abstractSchematic representation of a fluorometric method for determination of the CCAAT/enhancer binding protein alpha (C/EBPα). Fluorescently labeled dsDNA that can bind C/EBPα is introduced as a fluorescent probes. The dsDNA is electrostatically adsorbed on the positively charged AuNPs to quench their fluorescence. In the presence of C/EBPα, it will bind dsDNA which then diffuses away. The fluorescence of the AuNPs becomes restored.
一种用于测定 CCAAT/增强子结合蛋白α（C/EBPα）的方法，C/EBPα 是脂肪细胞分化的调节剂。该方法基于 Cy3 标记的 DNA 如果被吸附在带正电荷的金纳米粒子（AuNPs）上，其红色荧光（激发/发射最大值为 548/562nm）会被猝灭。将可以结合 C/EBPα 的荧光标记双链 DNA 作为荧光探针引入。双链 DNA 通过静电吸附在带正电荷的 AuNPs 上使其荧光猝灭。在存在 C/EBPα 的情况下，它将结合双链 DNA，然后扩散开来。AuNPs 的荧光得到恢复。荧光信号在 0.05 至 600ng·mL μM C/EBPα 浓度范围内呈线性增加，检测限为 29pg·mL。该方法具有特异性，并应用于分析细胞裂解物和原位。荧光测定 CCAAT/增强子结合蛋白α（C/EBPα）的示意图。将可以结合 C/EBPα 的荧光标记双链 DNA 作为荧光探针引入。双链 DNA 通过静电吸附在带正电荷的 AuNPs 上使其荧光猝灭。在存在 C/EBPα 的情况下，它将结合双链 DNA，然后扩散开来。AuNPs 的荧光得到恢复。
