采用凝胶内激酶分析评估植物中的激酶活性。
Assessing Kinase Activity in Plants with In-Gel Kinase Assays.
作者信息
Wang Pengcheng, Zhu Jian-Kang
机构信息
Department of Horticulture and Landscape Architecture, Purdue University, 625 Agriculture Mall Drive, West Lafayette, IN, 47907, USA.
Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences, Shanghai Institutes for Biological Sciences, Shanghai, 200032, China.
出版信息
Methods Mol Biol. 2016;1363:189-97. doi: 10.1007/978-1-4939-3115-6_15.
Abstract
The in-gel protein kinase assay is a powerful method to measure the protein phosphorylation activity of specific protein kinases. Any protein substrate can be embedded in polyacrylamide gels where they can be phosphorylated by protein kinases that are separated in the gel under denaturing conditions and then renatured. The kinase activity can be visualized in situ in the gels by autoradiography. This method has been used to compare the activities of protein kinases in parallel samples or to identify their potential substrates. Here, we describe in detail an in-gel kinase assay to measure the activity of some protein kinases in plants.
摘要
凝胶内蛋白激酶测定法是一种用于测量特定蛋白激酶的蛋白磷酸化活性的强大方法。任何蛋白底物都可以包埋在聚丙烯酰胺凝胶中，在变性条件下，凝胶中的蛋白激酶可使其磷酸化，然后再复性。通过放射自显影可在凝胶中原位观察激酶活性。该方法已用于比较平行样品中蛋白激酶的活性或鉴定其潜在底物。在此，我们详细描述一种用于测量植物中某些蛋白激酶活性的凝胶内激酶测定法。
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