GST-Perfringolysin O production for the localization and quantification of membrane cholesterol in human and mouse brain and liver

STAR Protocols

Fernanda Castro¹  Susana Nuñez²  Fabian Arenas³  Carmen Garcia-Ruiz³  Leire Goicoechea⁴  José C. Fernandez-Checa⁴  Sandra Torres⁴ 

[1] Centro de Investigación Biomédica en Red (CIBEREHD), Barcelona, Spain;Corresponding author;Liver Unit, Hospital Clinic i Provincial de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain;Department of Cell Death and Proliferation, Institute of Biomedical Research of Barcelona (IIBB), CSIC, Barcelona, Spain;

关键词: Cell Biology;    Cell Membrane;    Health Sciences;    Metabolism;    Microscopy;    Model Organisms;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: Abnormal cholesterol metabolism is linked to many neurodegenerative disorders. Here, we present a protocol for the production of a recombinant protein consisting of a Glutathione-S-Transferase tag fused with the Perfringolysin O (PFO). The GST-PFO tag enables analysis of the localization of cholesterol in subcellular membranes of human and mice brain and liver tissues. We have used this approach for samples from Niemann-Pick type C disease and non-alcoholic steatohepatitis models. The construct may also have applications for the diagnosis of cholesterol-accumulating disorders.For complete details on the use and execution of this protocol, please refer to Kwiatkowska et al. (2014).

【 授权许可】

Unknown