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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

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: 9 October 2023
Revised Date: 29 December 2023

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.
- Non-site-specific fluorescent labeling,
- Protein,
- Fluorescein isothiocyanate,
- Chemical biology experiment
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