Xi Wang , Maowei Ni , Chao Niu , Xinliang Zhu , Ting Zhao , Zhongxin Zhu , Yuanhu Xuan , Weitao Cong
{"title":"槲皮素对SDS-PAGE中磷酸化蛋白的简单检测","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":"{\"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}","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}
Simple detection of phosphoproteins in SDS-PAGE by quercetin
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.
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