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Updated: May 13, 2025
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Published on: December 30, 2016
Protocol for in vitro phase separation of N terminus of CIDEC proteins
Yiming Lu1, Zanzan Zhu1, Jianqin Wang2
1Shanghai Key Laboratory of Metabolic Remodeling and Health, State Key Laboratory of Genetics and Development of Complex Phenotypes, Institute of Metabolism and Integrative Biology, School of Life Sciences, Department of Endocrinology and Metabolism, Zhongshan Hospital, Fudan University, Shanghai 200438, China.
Abstract:
Cell death-inducing DFF45-like effector C (CIDEC) condensation through N-terminal phase separation is a prerequisite for CIDEC-mediated lipid droplet (LD) fusion and lipid storage promotion. Here, we present a protocol for in vitro phase separation of the N terminus of CIDEC (CIDEC-N) proteins. We describe steps for plasmid construction, protein expression, and purification. We then detail procedures for concentration detection and in vitro phase separation. This protocol provides a framework to investigate CIDEC-N phase separation properties, laying the foundation for further exploration of its role in lipid metabolism and obesity. For complete details on the use and execution of this protocol, please refer to Lyu et al.1.
