Stasyk Taras, Huber Lukas Alfons
Division of Cell Biology, Biocenter, Innsbruck Medical University, Innrain 80-82, A-6020, Innsbruck, Austria.
Methods Mol Biol. 2018;1664:79-86. doi: 10.1007/978-1-4939-7268-5_7.
Here, we describe the detailed step-by-step protocol for detection of phosphoproteins in two-dimensional difference gel electrophoresis (DIGE) gels. A standard DIGE protocol is combined with subsequent post-staining with phosphospecific fluorescent dye. The combination of these two methods complements DIGE-based proteome profiling by fluorescence detection of phosphoproteins in the same gel providing additional possibility for sensitive and accurate quantification of the differentially regulated phosphoproteins in biological samples.
在此，我们描述了在二维差异凝胶电泳（DIGE）凝胶中检测磷酸化蛋白的详细分步方案。将标准的DIGE方案与随后用磷酸特异性荧光染料进行的染色后处理相结合。这两种方法的结合通过在同一凝胶中对磷酸化蛋白进行荧光检测，对基于DIGE的蛋白质组分析进行补充，为生物样品中差异调节的磷酸化蛋白的灵敏和准确定量提供了额外的可能性。
