Emiko Kinoshita-Kikuta, K. Akayama, E. Kinoshita, T. Koike
{"title":"A dot-blot-staining method for detecting phosphoproteins with a Phos-tag Aqua fluorescent dye","authors":"Emiko Kinoshita-Kikuta, K. Akayama, E. Kinoshita, T. Koike","doi":"10.2198/jelectroph.64.7","DOIUrl":null,"url":null,"abstract":"We describe a method for detecting phosphoproteins by dot-blot staining with a Phos-tag Aqua fluorescent dye. By using the method, three types of in vitro protein kinase assays for visualizing Tyr-, His-, and Asp-phosphoproteins, respectively, were performed. The staining procedure, which requires less than 2.5 h to complete, is conducted under conditions of neutral pH that are particularly advantageous for detecting labile Hisand Asp-phosphoproteins. This method promises rapid and easy detection of phosphoproteins, and should be useful in high-throughput profiling of in vitro kinase activities or in vitro kinase inhibition.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"111 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of capillary electrophoresis","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.2198/jelectroph.64.7","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
We describe a method for detecting phosphoproteins by dot-blot staining with a Phos-tag Aqua fluorescent dye. By using the method, three types of in vitro protein kinase assays for visualizing Tyr-, His-, and Asp-phosphoproteins, respectively, were performed. The staining procedure, which requires less than 2.5 h to complete, is conducted under conditions of neutral pH that are particularly advantageous for detecting labile Hisand Asp-phosphoproteins. This method promises rapid and easy detection of phosphoproteins, and should be useful in high-throughput profiling of in vitro kinase activities or in vitro kinase inhibition.