Kamada Yuka, Fukuda Ryosuke, Okiyoneda Tsukasa
Department of Biomedical Chemistry, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo 669-1337, Japan.
Bio Protoc. 2019 Nov 20;9(22):e3430. doi: 10.21769/BioProtoc.3430.
Ubiquitylation is a common post-translational modification of cellular proteins that results in proteasomal and lysosomal degradations. Ubiquitylation is generally measured by methods such as immunoblotting using anti-ubiquitin antibodies after isolating the protein-of-interest by denaturing immunoprecipitation. The following protocol can be used to easily quantify the ubiquitylation of the protein-of-interest tagged with biotin by ELISA.
泛素化是细胞蛋白质常见的翻译后修饰，可导致蛋白酶体和溶酶体降解。泛素化通常通过以下方法测量：在通过变性免疫沉淀分离出目标蛋白后，使用抗泛素抗体进行免疫印迹。以下方案可用于通过酶联免疫吸附测定（ELISA）轻松定量生物素标记的目标蛋白的泛素化。
