1. 05SAR-PAGE: Separation of protein dimerization and modification using a gel with 0.05% sarkosyl.
- Author
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Huang, Liqun, Kou, Xinhui, Zheng, Wenwen, Xiao, Xiong, Li, Conggang, Liu, Maili, Liu, Yixiang, and Jiang, Ling
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POLYACRYLAMIDE gel electrophoresis , *PROTEIN fractionation , *CHEMICAL modification of proteins , *PROTEIN structure , *NUCLEAR magnetic resonance spectroscopy , *GEL electrophoresis - Abstract
A protein gel electrophoresis procedure using 0.05% w/v sarkosyl, is reported. The method called 05SAR-PAGE can be used to identify the native masses, dimeric states and modification states of proteins, and also be suitable for pursuing native electroblotting and immunodetection. It has been demonstrated by NMR spectroscopy that 0.05% w/v SAR is much milder than SDS, so it has subtle effects on the native structure of proteins. Therefore, the non-covalent dimerization of PhoB N and PhoR cp can be identified by 05SAR-PAGE which cannot be observed by SDS-PAGE. It has also been demonstrated that 05SAR-PAGE can be used to identify the phosphorylated or methylated proteins. Besides, 05SAR-PAGE shows the advantages of simple operation and low cost, and can be easily adapted to diverse applications. Image 1 • Determination of the dimerization states of PhoB N and PhoR cp by 05SAR-PAGE. • Mild effect of SAR on protein structure. • Western-blot of monomer and dimer states in vivo by 05SAR-PAGE. • Identification of different modification states of proteins by 05SAR-PAGE. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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