植物蛋白的体外泛素化检测方法
赵庆臻1,刘利静2,谢旗3,4,于菲菲3,*(
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In Vitro Ubiquitination Assay for Plant Proteins
Qingzhen Zhao1,Lijing Liu2,Qi Xie3,4,Feifei Yu3,*()
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图2. SDIR1的自身泛素化反应(E3活性检测) (
在小麦E1、E2 (UBCH5b)和His-Ub存在的情况下, 检测MBP-SDIR1及其突变形式MBP-SDIR1 (H234Y)的E3活性, 以MBP蛋白作为阴性对照。反应产物经8% SDS-PAGE胶分离后, 分别用Nickel-HRP (左图)和anti-MBP抗体(右图)进行western blot检测。
Figure 2. Autoubiquitination assay (E3 activity assay) of SDIR1 (
The E3 ligase activity of MBP-SDIR1 and its mutation form MBP-SDIR1 (H234Y) was detected at the presence of E1, E2 (UBCH5b) and His-Ub. MBP acts as the negative control. The reaction products were resolved by 8% SDS-PAGE gel and then western blot was performed with Nickel-HRP (left part) or anti-MBP (right part) antibody, respectively.
