Rao Chethana, Khan Syamantak, Verma Navneet C, Nandi Chayan Kanti
School of Basic Sciences, Indian Institute of Technology, Mandi, Himachal Pradesh, 175001, India.
Chembiochem. 2017 Dec 14;18(24):2385-2389. doi: 10.1002/cbic.201700440. Epub 2017 Nov 2.
We present efficient labelling of several proteins with orange-emissive carbon dots. N-Hydroxysuccinimide was used to activate the carboxyl groups of carbon dots, which subsequently reacted with the lysine groups present on the protein. Labelling was confirmed by UV absorption spectroscopy, PAGE and fluorescence correlation spectroscopy. Protein-conjugated carbon dots showed an enhancement in fluorescence lifetime and intensity owing to reduced intramolecular dynamic fluctuations. Single-molecule fluorescence measurements showed reduced fluorescence fluctuations and higher photon budget after protein tagging. Our study opens up opportunities to use carbon dots as highly precise biolabelling probes.
我们展示了用橙色发射碳点对几种蛋白质进行高效标记。使用N-羟基琥珀酰亚胺活化碳点的羧基,其随后与蛋白质上存在的赖氨酸基团反应。通过紫外吸收光谱、聚丙烯酰胺凝胶电泳和荧光相关光谱对标记进行了确认。由于分子内动态波动减少,蛋白质共轭碳点的荧光寿命和强度有所增强。单分子荧光测量表明,蛋白质标记后荧光波动减少,光子预算更高。我们的研究为将碳点用作高精度生物标记探针开辟了机会。