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Updated: Jan 28, 2026

Generation, Purification, and Characterization of Cell-invasive DISC1 Protein Species
Published on: August 30, 2012
Purification and Characterization of Human Niemann-Pick C1 Protein
Xin Gong1,2, Hongwu Qian3
1Department of Molecular Biology, Princeton University, Princeton, NJ, USA. gongx@sustc.edu.cn.
Abstract:
Niemann-Pick C1 (NPC1) is a membrane protein required for the transport of low-density lipoprotein (LDL)-derived cholesterol from endosomes and lysosomes to the other organelles. Here, we describe the recombinant protein expression, purification, and characterization of the human NPC1. The protein is transiently expressed in human embryonic kidney (HEK) cells. Our purification protocol describes the steps to obtain a pure and homogeneous NPC1 protein. Niemann-Pick C2 (NPC2) is a small soluble protein, which mediates cholesterol transport in tandem with NPC1. Finally, we also describe two biochemical approaches to characterize NPC1 function in vitro-a cholesterol transfer assay from purified NPC2 to NPC1 and a binding assay between NPC1 and NPC2.
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