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

Ying Miao, Qian Du, Hong Zhang, Yukang Yuan, Yibo Zuo, Hui Zheng

2023BIO-PROTOCOL60 citationsDOIOpen Access PDF

Abstract

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.

Topics & Concepts

CycloheximideProtein biosynthesisIntracellularProtein degradationProtein synthesis inhibitorRibosomeCell biologyBiochemistryProteasomeTranslation (biology)ChemistryBiologyRNAMessenger RNAGeneUbiquitin and proteasome pathwaysProtein Degradation and InhibitorsPeptidase Inhibition and Analysis
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