蛋白质的非定点荧光标记——推荐一个化学生物学实验

引用本文: 洪歆怡, 薛太玲, 徐洲, 谢恩蓉, 吴明凯, 王青青, 吴丽娜. 蛋白质的非定点荧光标记——推荐一个化学生物学实验[J]. 大学化学, 2024, 39(4): 351-360. doi: 10.3866/PKU.DXHX202310010 shu

Citation: Xinyi Hong, Tailing Xue, Zhou Xu, Enrong Xie, Mingkai Wu, Qingqing Wang, Lina Wu. Non-Site-Specific Fluorescent Labeling of Proteins as a Chemical Biology Experiment[J]. University Chemistry, 2024, 39(4): 351-360. doi: 10.3866/PKU.DXHX202310010 shu

蛋白质的非定点荧光标记——推荐一个化学生物学实验

厦门大学化学化工学院, 化学国家级实验教学示范中心(厦门大学), 福建厦门 361005

通讯作者: 吴丽娜, alina1222@xmu.edu.cn

基金项目:
2020年厦门大学教学改革研究项目

摘要: 本论文介绍了一项化学生物学实验，旨在利用异硫氰酸荧光素的异硫氰基与蛋白质氨基共价结合的原理，实现对体外蛋白(以牛血清白蛋白为例)和体内蛋白(以大肠杆菌总蛋白为例)的非定点荧光标记。实验采用凝胶过滤法作为后续纯化手段，并运用紫外-可见分光光度计、流式细胞术以及荧光显微镜对标记结果进行全面表征。该实验涵盖了多种基础和前沿实验技能，与科研工作中常见的抗体荧光标记技术密切相关，难度适中。通过参与这一实验，学生不仅能够深入理解化学生物学基础知识，提高综合实践能力，还能够拓宽科研视野并培养科研思维。

关键词:

English

Non-Site-Specific Fluorescent Labeling of Proteins as a Chemical Biology Experiment

National Demonstration Center for Experimental Chemistry Education (Xiamen University), College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, Fujian Province, China

Corresponding author: Lina Wu, alina1222@xmu.edu.cn

Received Date: 09 October 2023
Revised Date: 29 December 2023
Available Online: 09 January 2024

Abstract: This paper presents a chemical biology experiment designed to exploit the principle of covalent binding between fluorescein isothiocyanate and protein amines. This process is aimed at achieving non-site-specific fluorescent labeling of both extracellular proteins, exemplified by bovine serum albumin, and intracellular proteins, using Escherichia coli total protein as a case study. Gel filtration chromatography is employed as a subsequent purification method, with comprehensive characterization of the labeling results performed using UV-Visible spectrophotometry, flow cytometry, and fluorescence microscopy. This experiment encompasses a range of foundational and cutting-edge laboratory skills. It is closely aligned with the commonly employed antibody fluorescence labeling techniques in scientific research, presenting a moderate level of complexity. Participation in this experiment allows students to gain a profound understanding of fundamental concepts in chemical biology, enhance their comprehensive practical skills, broaden their research perspectives and nurture scientific thinking.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

蛋白质的非定点荧光标记——推荐一个化学生物学实验

通讯作者: 吴丽娜, alina1222@xmu.edu.cn

English

Non-Site-Specific Fluorescent Labeling of Proteins as a Chemical Biology Experiment

Corresponding author: Lina Wu, alina1222@xmu.edu.cn

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

蛋白质的非定点荧光标记——推荐一个化学生物学实验

通讯作者: 吴丽娜, alina1222@xmu.edu.cn

English

Non-Site-Specific Fluorescent Labeling of Proteins as a Chemical Biology Experiment

Corresponding author: Lina Wu, alina1222@xmu.edu.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

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