Nagy Zoltan, Comer Shane, Smolenski Albert
Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, United Kingdom.
UCD School of Medicine, University College Dublin, Belfield, Dublin, Ireland.
Curr Protoc Protein Sci. 2018 Aug;93(1):e64. doi: 10.1002/cpps.64. Epub 2018 Jul 25.
Phos-tag gels are recent tools to dissect protein phosphorylation that operate by inducing a shift in the electrophoretic mobility of phosphorylated proteins compared to their nonphosphorylated counterparts. This article describes the preparation and electrophoresis of Zn -Phos-tag gels along with electrotransfer of the separated phospho- and nonphosphoproteins onto a PVDF membrane using either wet-tank or semidry transfer. We also discuss the theory behind the technology with critical parameters to keep in mind for its successful application. © 2018 by John Wiley & Sons, Inc.
Phos-tag凝胶是用于分析蛋白质磷酸化的最新工具，其作用原理是与未磷酸化的对应蛋白质相比，诱导磷酸化蛋白质的电泳迁移率发生变化。本文介绍了Zn-Phos-tag凝胶的制备和电泳，以及使用湿槽转移或半干转移将分离的磷酸化和非磷酸化蛋白质电转移到PVDF膜上的方法。我们还讨论了该技术背后的理论以及成功应用该技术时需要牢记的关键参数。© 2018约翰威立父子公司。
