05SAR-PAGE：使用含 0.05%Sarkosyl 的凝胶分离蛋白质二聚化和修饰。

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

机构信息

Key Laboratory of Magnetic Resonance in Biological System, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Wuhan Center for Magnetic Resonance, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan, 430071, China; University of Chinese Academy of Sciences, Beijing, 100049, China.

出版信息

Anal Chim Acta. 2020 Mar 8;1101:193-198. doi: 10.1016/j.aca.2019.12.013. Epub 2019 Dec 9.

DOI:10.1016/j.aca.2019.12.013

PMID:32029111

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 and PhoR 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.

摘要

本文报道了一种使用 0.05%w/v 十二烷基肌氨酸钠（sarkosyl）的蛋白质凝胶电泳程序。该方法称为 05SAR-PAGE，可用于鉴定蛋白质的天然分子量、二聚体状态和修饰状态，也适用于进行天然电印迹和免疫检测。通过 NMR 光谱证明，0.05%w/v SAR 比 SDS 温和得多，因此对蛋白质的天然结构影响很小。因此，通过 05SAR-PAGE 可以鉴定 PhoB 和 PhoR 的非共价二聚体，而 SDS-PAGE 则无法观察到这一点。也已证明 05SAR-PAGE 可用于鉴定磷酸化或甲基化蛋白质。此外，05SAR-PAGE 具有操作简单、成本低的优点，并且易于适应各种应用。