A nile red fluorescence assay for LC3 autophagy protein binding to lipid bilayers.

ABSTRACT: Autophagy consists of the degradation and recycling of defective or aging cellular components. This process occurs in all eukaryotes, and starts by the formation of a double-bilayer structure known as the autophagosome. In humans, autophagosome generation requires, among others, the action of three homologous proteins, designated as LC3A, LC3B, and LC3C. These are amphipathic proteins, which can exist either in aqueous or in membranous environments. Quantification of LC3 binding to lipid bilayers is usually achieved by a rather cumbersome procedure, involving separation of bound and free forms by density gradient centrifugation and fractional analysis of the centrifuged samples. This paper describes a simple protein binding assay based on the fluorescence properties of Nile red. This solva

SUBMITTER: Sot J 

PROVIDER: S-EPMC12494833 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature