Xi Wang , Maowei Ni , Chao Niu , Xinliang Zhu , Ting Zhao , Zhongxin Zhu , Yuanhu Xuan , Weitao Cong
{"title":"Simple detection of phosphoproteins in SDS-PAGE by quercetin","authors":"Xi Wang , Maowei Ni , Chao Niu , Xinliang Zhu , Ting Zhao , Zhongxin Zhu , Yuanhu Xuan , Weitao Cong","doi":"10.1016/j.euprot.2014.07.002","DOIUrl":null,"url":null,"abstract":"<div><p>A novel fluorescence-based staining method was developed for phosphoprotein analysis in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Similar to the mechanism of immobilized metal ion affinity chromatography (IMAC), the method employed quercetin–aluminum (III)-appended complex as a fluoroprobe to selectively visualize phosphorylated proteins among total proteins. According to the results, as low as 16–32<!--> <!-->ng of phosphoproteins (α-casein, β-casein and phosvitin) could be selectively detected in 90<!--> <!-->min with a wide linear dynamic range. In addition, the specificity of this novel stain for phosphoproteins was confirmed by 1-D and 2-D SDS-PAGE, dephosphorylation, western blot and liquid chromatography–mass spectrometry analysis (LC–MS/MS), respectively.</p></div>","PeriodicalId":38260,"journal":{"name":"EuPA Open Proteomics","volume":"4 ","pages":"Pages 156-164"},"PeriodicalIF":0.0000,"publicationDate":"2014-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.euprot.2014.07.002","citationCount":"9","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"EuPA Open Proteomics","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2212968514000476","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 9
Abstract
A novel fluorescence-based staining method was developed for phosphoprotein analysis in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Similar to the mechanism of immobilized metal ion affinity chromatography (IMAC), the method employed quercetin–aluminum (III)-appended complex as a fluoroprobe to selectively visualize phosphorylated proteins among total proteins. According to the results, as low as 16–32 ng of phosphoproteins (α-casein, β-casein and phosvitin) could be selectively detected in 90 min with a wide linear dynamic range. In addition, the specificity of this novel stain for phosphoproteins was confirmed by 1-D and 2-D SDS-PAGE, dephosphorylation, western blot and liquid chromatography–mass spectrometry analysis (LC–MS/MS), respectively.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
