Analysis of Protein Phosphorylation Using Phos-Tag Gels.
Analysis of Protein Phosphorylation Using Phos-Tag Gels.
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DOI：
10.1002/cpps.64
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发表时间：
2018-08-01
影响因子：
--
通讯作者：
Smolenski, Albert
中科院分区：
文献类型：
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作者：
Nagy, Zoltan;Comer, Shane;Smolenski, Albert
PHOS-TAG凝胶是最近用来分析蛋白质磷酸化的工具，它通过诱导磷酸化蛋白质的凝胶迁移率比非磷酸化蛋白质的凝胶迁移率发生变化来发挥作用。本文介绍了锌磷标签凝胶的制备和电泳法，以及用湿法或半干法将分离的磷酸和非磷酸蛋白电转移到PVDF膜上。我们还讨论了该技术背后的理论和关键参数，以记住它的成功应用。2018年John Wiley&Sons，Inc.
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 Zn2+ -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.