{"title":"A silver staining procedure for proteins in agarose gels","authors":"K. Yokomizo, S. Maruya, N. Hiratsuka, K. Shiba","doi":"10.2198/JELECTROPH.50.1","DOIUrl":null,"url":null,"abstract":"We modified the silver staining method of Kerenyi and Gallyas (Clin Chim Acta 1972; 38: 465-467) for visualizing low levels of protein with our own agarose gels as reported previously (Yokomizo K. et al. J Electrophoresis 2003; 47: 91-97). To reduce background staining and avoid the formation of artifact spots, a number of factors were studied. These included the composition of the staining reagent and fixative solutions (first and second fixation). It was found that 2.5% Na2CO3, 0.1% AgNO3, 0.1% NH4NO3, 0.75% tungtosilicic acid and 0.14% formaldehyde of the staining reagent mixture was critical and that the addition of 5% ZnSO4 to the second fixative solution resulted in consistently lower background staining and a significant reduction in artifact spots. Our silver staining procedure is approximately 100-fold more sensitive than Coomassie brilliant blue and the detection limit of bovine serum albumin is about 1.46 ng per band.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"18 1","pages":"1-5"},"PeriodicalIF":0.0000,"publicationDate":"2006-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of capillary electrophoresis","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.2198/JELECTROPH.50.1","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 1
Abstract
We modified the silver staining method of Kerenyi and Gallyas (Clin Chim Acta 1972; 38: 465-467) for visualizing low levels of protein with our own agarose gels as reported previously (Yokomizo K. et al. J Electrophoresis 2003; 47: 91-97). To reduce background staining and avoid the formation of artifact spots, a number of factors were studied. These included the composition of the staining reagent and fixative solutions (first and second fixation). It was found that 2.5% Na2CO3, 0.1% AgNO3, 0.1% NH4NO3, 0.75% tungtosilicic acid and 0.14% formaldehyde of the staining reagent mixture was critical and that the addition of 5% ZnSO4 to the second fixative solution resulted in consistently lower background staining and a significant reduction in artifact spots. Our silver staining procedure is approximately 100-fold more sensitive than Coomassie brilliant blue and the detection limit of bovine serum albumin is about 1.46 ng per band.