大学化学2024，Vol.39Issue(4)：351-360,10.DOI:10.3866/PKU.DXHX202310010
蛋白质的非定点荧光标记
Non-Site-Specific Fluorescent Labeling of Proteins as a Chemical Biology Experiment
摘要
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.关键词
非定点荧光标记/蛋白质/异硫氰酸荧光素/化学生物学实验Key words
Non-site-specific fluorescent labeling/Protein/Fluorescein isothiocyanate/Chemical biology experiment分类
社会科学引用本文复制引用
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[J].大学化学,2024,39(4):351-360,10.基金项目
2020年厦门大学教学改革研究项目 （）
