05SAR-PAGE: Separation of protein dimerization and modification using a gel with 0.05% sarkosyl

• Published in: Analytica chimica acta 2020-03, Vol.1101, p.193-198
• Main Authors: Huang, Liqun, Kou, Xinhui, Zheng, Wenwen, Xiao, Xiong, Li, Conggang, Liu, Maili, Liu, Yixiang, Jiang, Ling
• Format: Article
• Language: English
• Subjects: Dimerization; Gel electrophoresis; Immunodetection; Modification states; NMR spectroscopy; Sarkosyl
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2019.12.013

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 PhoBN and PhoRcp 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. [Display omitted] •Determination of the dimerization states of PhoBN and PhoRcp 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.