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