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Citation: Baoxin Zhang, Zhuo Fan, Chenxin Yu, Zhe Liu, Yongwen Shen, Xinping Hui, Jianguo Fang. 蛋白质中总巯基的测定及标记[J]. University Chemistry, ;2022, 37(5): 211107. doi: 10.3866/PKU.DXHX202111072 shu

蛋白质中总巯基的测定及标记

National Demonstration Center for Experimental Education, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China

Corresponding author: Jianguo Fang, fangjg@lzu.edu.cn
Received Date: 24 November 2021

生物硫醇的检测与蛋白质荧光标记是当今科学研究的热门领域。本实验使用传统的Ellman法及Naph-EA-Mal荧光探针标记法对牛血清白蛋白（Bovine Serum Albumin，BSA）的巯基进行检测，并利用该探针实现了细胞内巯基的原位标记，使学生对传统方法和前沿热点均有较好的掌握和了解。通过本实验的开展，学生不仅可以清楚了解蛋白三维结构对其所含基团的反应活性的影响，而且可以从化学层面理解硫醇类探针的工作原理及活细胞中生物硫醇的原位成像。实验内容涵盖蛋白质还原、变性、生物大分子与小分子的分离、生物大分子的化学修饰以及活细胞荧光成像等，综合性强且容易操作。教学内容可以实现模块化，便于不同学校根据条件选取相应的教学内容。教学实践不仅可以让学生复习巩固化学生物学知识，也帮助学生了解前沿交叉学科热点，同时在虚拟实验平台开设了补充实验，使得教学更加立体化。
- Protein sulfhydryl group,
- Ellman method,
- Fluorescent probe,
- Live cell imaging,
- Protein chemical modifications
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