Li Qingzhong, Yuan Xianping, Zhao Yang
Key Laboratory of Plant Carbon Capture, Shanghai Center for Plant Stress Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
Key Laboratory of Plant Carbon Capture, Shanghai Center for Plant Stress Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai 200032, China; University of Chinese Academy of Sciences, Beijing 100049, China.
STAR Protoc. 2025 Jun 20;6(2):103842. doi: 10.1016/j.xpro.2025.103842. Epub 2025 May 28.
The SNF1-regulated protein kinase 2s (SnRK2s) are activated by phytohormone abscisic acid (ABA) and osmotic stress to control plant growth and stress responses; however, assessing SnRK2 activity is challenging. Here, we present a protocol to detect SnRK2 activity in plants. We describe steps for performing immunoblotting with anti-phospho-S175-SnRK2 antibody, in-gel kinase assay, band-shift assay, and immunoprecipitated kinase assay. Immunoblotting and in-gel kinase assays are suitable for evaluating endogenous SnRK2 activity, whereas band-shift and immunoprecipitated kinase assays are applicable to assess tagged SnRK2 activity in transgenic lines. For complete details on the use and execution of this protocol, please refer to Li et al., Li et al., and Yuan et al..
蔗糖非发酵1相关蛋白激酶2（SnRK2）受植物激素脱落酸（ABA）和渗透胁迫激活，以控制植物生长和胁迫反应；然而，评估SnRK2活性具有挑战性。在此，我们提供了一种在植物中检测SnRK2活性的方法。我们描述了用抗磷酸化-S175-SnRK2抗体进行免疫印迹、凝胶内激酶测定、条带迁移测定和免疫沉淀激酶测定的步骤。免疫印迹和凝胶内激酶测定适用于评估内源性SnRK2活性，而条带迁移和免疫沉淀激酶测定适用于评估转基因系中标记的SnRK2活性。有关本方法使用和执行的完整详细信息，请参考李等人、李等人和袁等人的文献。
