Chakraborty Anirban, Wang Dongye, Ebright Yon W, Ebright Richard H
Department of Chemistry and Chemical Biology, Waksman Institute, Rutgers University, Piscataway, New Jersey, USA.
Methods Enzymol. 2010;472:19-30. doi: 10.1016/S0076-6879(10)72018-8.
We describe the synthesis of phosphine derivatives of three fluorescent probes that have a brightness and photostability suitable for single-molecule fluorescence spectroscopy and microscopy: Alexa488, Cy3B, and Alexa647. In addition, we describe procedures for use of these reagents in azide-specific, bioorthogonal labeling through Staudinger-Bertozzi ligation, as well as procedures for the quantitation of labeling specificity and labeling efficiency. The reagents and procedures of this report enable chemoselective, site-selective labeling of azide-containing biomolecules for single-molecule fluorescence spectroscopy and microscopy.
我们描述了三种荧光探针的膦衍生物的合成，这些探针具有适合单分子荧光光谱和显微镜的亮度和光稳定性：Alexa488、Cy3B和Alexa647。此外，我们还描述了通过施陶丁格-贝托齐连接在叠氮化物特异性生物正交标记中使用这些试剂的程序，以及标记特异性和标记效率的定量程序。本报告中的试剂和程序能够对含叠氮化物的生物分子进行化学选择性、位点选择性标记，用于单分子荧光光谱和显微镜。
