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
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通过 4',5'-二溴荧光素在 SDS-PAGE 中检测阴性蛋白质的超灵敏染色剂
DOI：
10.1016/j.jprot.2017.06.014
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发表时间：
2017
影响因子：
3.3
通讯作者：
Zhu Zhongxin
中科院分区：
文献类型：
--
作者：
Yu Dongdong;Wang Yang;Zhang Shaoqing;Chen Zhiwei;Xue Mei;Wang Ying;Cong Weitao;Jin Litai;Zhu Zhongxin
建立了一种简便、经济、灵敏的4′，5 ′-二溴荧光素（DBF）染色十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）蛋白质的方法。DBF染色法仅需2步即可在10 min内检测到0.025-0.05 ng蛋白，比Eosin Y（EY）和SYPRO Ruby染色法敏感10倍，比咪唑锌（IZ）阴性染色法敏感20倍。此外，LC-MS/MS结果表明，新开发的染色方法与下游蛋白质鉴定兼容。并通过分子对接的方法探讨了DBF染色的机理。
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 0.025–0.05 ng protein could be detected within 10 min (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.