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Updated: May 12, 2026

Protein Misfolding Cyclic Amplification of Prions
Published on: November 7, 2012
Protocol for investigating phase transition of prion-like proteins in vitro with concentration variation
Xiao-Tong Chen1, Yu-Meng Sun1, Wen-Tao Wang1
1MOE Key Laboratory of Gene Function and Regulation, Guangdong Province Key Laboratory of Pharmaceutical Functional Genes, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
Abstract:
Protein concentration is an important factor regulating liquid-to-solid phase transition of proteins with prion-like domains (PLDs). Here, we present a protocol for investigating the effect of prion-like protein concentration on phase transition of fibrillarin (FBL) condensates in vitro using phase separation assays. We describe steps for construction of fluorescent-protein-tagged FBL plasmids, protein expression and purification in vitro, and detection of condensed states of FBL. This protocol has potential applications in phase separation assays for studying phase transitions in any prion-like protein. For complete details on the use and execution of this protocol, please refer to Sun et al.1.
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