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{"title":"Protein Detection in Gels Using Fixation.","authors":"Lynn A Beer, David W Speicher","doi":"10.1002/cpps.48","DOIUrl":null,"url":null,"abstract":"<p><p>The most commonly used types of gels for separating proteins are SDS gels, either in a 1-D format or as the second dimension of various 2-D separations, and the most common methods of visualizing proteins in these gels use protein binding dyes after fixing the proteins in the gel matrix. In recent years, there has been a continuing trend away from preparing staining solutions in the laboratory to using commercially available kits, which are convenient, save time, have defined shelf lives, and may provide greater reproducibility than stains formulated in research laboratories. In general, when using commercial kits, satisfactory results can be readily obtained by following the manufacturer's protocols. This unit reviews commonly used fixation-based stains and provides a number of manual formulations with staining protocols for those who prefer such staining methods. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":10866,"journal":{"name":"Current Protocols in Protein Science","volume":"91 ","pages":"10.5.1-10.5.20"},"PeriodicalIF":0.0000,"publicationDate":"2018-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpps.48","citationCount":"21","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Protein Science","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1002/cpps.48","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
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Abstract
The most commonly used types of gels for separating proteins are SDS gels, either in a 1-D format or as the second dimension of various 2-D separations, and the most common methods of visualizing proteins in these gels use protein binding dyes after fixing the proteins in the gel matrix. In recent years, there has been a continuing trend away from preparing staining solutions in the laboratory to using commercially available kits, which are convenient, save time, have defined shelf lives, and may provide greater reproducibility than stains formulated in research laboratories. In general, when using commercial kits, satisfactory results can be readily obtained by following the manufacturer's protocols. This unit reviews commonly used fixation-based stains and provides a number of manual formulations with staining protocols for those who prefer such staining methods. © 2018 by John Wiley & Sons, Inc.
用固定法检测凝胶中的蛋白质。
用于分离蛋白质的最常用的凝胶类型是SDS凝胶,无论是以一维格式还是作为各种二维分离的第二维,在这些凝胶中可视化蛋白质的最常用方法是在将蛋白质固定在凝胶基质中后使用蛋白质结合染料。近年来,有一种持续的趋势,从在实验室准备染色溶液到使用市售试剂盒,这些试剂盒方便,节省时间,有明确的保质期,并且可能提供比研究实验室配制的染色剂更大的可重复性。一般来说,当使用商业试剂盒时,遵循制造商的协议可以很容易地获得满意的结果。本单元回顾了常用的基于固定的染色,并为那些喜欢这种染色方法的人提供了许多带有染色方案的手工配方。©2018 by John Wiley & Sons, Inc。
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