2. Academic Validation
  3. An ultrasensitive stain for negative protein detection in SDS-PAGE via 4',5'-Dibromofluorescein

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

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: jin_litai@126.com.
  • 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: zhongxinzhu2010@163.com.
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.

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