Cycloheximide (CHX) Chase Assay to Examine Protein Half-life

Ying Miao; Qian Du; Hong-Guang Zhang; Yukang Yuan; Yibo Zuo; Hui Zheng

doi:10.21769/BioProtoc.4690

Cycloheximide (CHX) Chase Assay to Examine Protein Half-life

Bio Protoc. 2023 Jun 5;13(11):e4690. doi: 10.21769/BioProtoc.4690.

Authors

Ying Miao^{1

2}, Qian Du^{1

2}, Hong-Guang Zhang^{1

2}, Yukang Yuan^{1

2}, Yibo Zuo^{1

2}, Hui Zheng^{1

2}

Affiliations

¹ Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, China.
² Jiangsu Key Laboratory of Infection and Immunity, Soochow University, Suzhou 215123, China.

Abstract

Cycloheximide (CHX) is a small molecule derived from Streptomyces griseus that acts as fungicide. As a ribosome inhibitor, CHX can restrict the translation elongation of eukaryotic protein synthesis. Once protein synthesis is inhibited by CHX, the level of intracellular proteins decreases by degradation through the proteasome or lysosome system. Thus, the CHX chase assay is widely recognized and used to observe intracellular protein degradation and to determine the half-life of a given protein in eukaryotes. Here, we present a complete experimental procedure of the CHX chase assay. Graphical overview.

Keywords: Cycloheximide (CHX) chase assay; Protein degradation; Protein half-life; Ubiquitin; Ubiquitination.