A method for rapid and sensitive negative staining of proteins in SDS-PAGE using 2',7'-dichlorofluorescein.

Yuan Zhang, Dongdong Yu, Jiangbo Dai, Xu Wang, Zhongxin Zhu, Yewei Huang, Jia Sun, Zhiwei Chen, Litai Jin, Yang Wang

Proteomics 2017 Jun

filter terms:

We report here a rapid and sensitive technique for negative visualization of protein in 1D and 2D SDS-PAGE by using 2', 7'-dichlorofluorescein (DCF), which appeared as transparent and colorless bands in an opaque gel matrix background. For DCF stain, down to 0.1-0.2 ng protein could be easily visualized within 7 min by only two steps, and the staining is fourfold more sensitive than that of Eosin Y (EY) negative stain and glutaraldehyde (GA) silver stain, and eightfold more sensitive than that of the commonly used imidazole-zinc (IZ) negative stain. Furthermore, DCF stain provided good reproducibility, linearity, and MS compatibility compared with those of IZ stain. In addition, the potential staining mechanism was investigated by colorimetric experiment and molecular docking, and the results demonstrated that the interaction between DCF and protein occurs mainly via van der waals force, electrostatic interaction, and hydrogen bonding. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Citation

Yuan Zhang, Dongdong Yu, Jiangbo Dai, Xu Wang, Zhongxin Zhu, Yewei Huang, Jia Sun, Zhiwei Chen, Litai Jin, Yang Wang. A method for rapid and sensitive negative staining of proteins in SDS-PAGE using 2',7'-dichlorofluorescein. Proteomics. 2017 Jun;17(12)