An ultrasensitive stain for negative protein detection in SDS-PAGE via 4',5'-Dibromofluorescein

  • J Proteomics. 2017 Aug 8;165:21-25. doi: 10.1016/j.jprot.2017.06.014.
Dongdong Yu  1 ,  Yang Wang  1 ,  Shaoqing Zhang  1 ,  Zhiwei Chen  1 ,  Mei Xue  1 ,  Ying Wang  1 ,  Weitao Cong  2 ,  Litai Jin  3 ,  Zhongxin Zhu  4
Affiliations
  • 1. Key Laboratory of Biotechnology Pharmaceutical Engineering of Zhejiang Province, School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325000, PR China.
  • 2. Key Laboratory of Biotechnology Pharmaceutical Engineering of Zhejiang Province, School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325000, PR China; Wenzhou Undersun Biotechnology Co., Ltd., Wenzhou 325000, PR China.
  • 3. Key Laboratory of Biotechnology Pharmaceutical Engineering of Zhejiang Province, School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325000, PR China; Wenzhou Undersun Biotechnology Co., Ltd., Wenzhou 325000, PR China. Electronic address: [email protected].
  • 4. Key Laboratory of Biotechnology Pharmaceutical Engineering of Zhejiang Province, School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325000, PR China; Wenzhou Undersun Biotechnology Co., Ltd., Wenzhou 325000, PR China. Electronic address: [email protected].
PMID: 28634119 DOI: 10.1016/j.jprot.2017.06.014
Abstract

A highly sensitive method for brief and economical staining of proteins in 1-D and 2-D sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) by 4',5'-Dibromofluorescein (DBF) was developed in this study. Down to 0.025-0.05ng protein could be detected within 10min (only 2 steps) by DBF stain, which is approximately 10-fold more sensitive than those of Eosin Y (EY) and SYPRO Ruby stains, and 20-fold more sensitive than that of imidazole-zinc (IZ) negative stain. In addition, the LC-MS/MS results indicated that the newly developed staining method is compatible with the downstream protein identification. Moreover, the mechanism of DBF stain was investigated by molecular docking.

Keywords
4′,5′-Dibromofluorescein; Negative stain; Protein detection; Proteomics; SDS-PAGE.
Products