Abstract:

Biological sulfhydryl detection and protein labeling are critical in current scientific research. In this study, the traditional Ellman method and the newly developed Naph-EA-Mal fluorescent probe were used to quantify sulfhydryl groups in bovine serum albumin, thus giving students a better understanding of both traditional methods and cutting-edge technology. The content of sulfhydryl can be detected in situ through measuring the fluorescence intensity. In addition, students gain knowledge regarding the mechanism of thiol probes and the influence of the protein's three-dimensional structure on the reactivity of the sulfhydryl groups within the protein. The experiment covers basic technology of treating cells and different methods of treating biomacromolecules. These methods are comprehensive, modularized, and easy to implement. This lab course will help students obtain cutting-edge chemical biology knowledge. In addition, the experiments on the virtual experiment platform could further help students learn and understand the principles and operational methods used in this experiment.

Key words: Protein sulfhydryl group, Ellman method, Fluorescent probe, Live cell imaging, Protein chemical modifications