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Updated: May 10, 2026
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A Quantitative Fluorescence Microscopy-based Single Liposome Assay for Detecting the Compositional Inhomogeneity Between Individual Liposomes
Published on: December 13, 2019
Protocol to determine in vitro lipid-protein interactions using liposome flotation assay
Karan Khadayat1, Morgan House1, Vasudeva Tati1
1Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37916, USA.
STAR Protocols
|May 8, 2026
Summary
This study details a gradient centrifugation method to analyze lipid-protein interactions. The protocol uses liposomes and purified proteins to understand cellular processes like metabolism and signaling.
Area of Science:
- Biochemistry
- Cell Biology
- Biophysics
Background:
- Lipid-protein interactions are fundamental to cellular functions, including metabolism, transport, and signaling pathways.
- Understanding these interactions is key to deciphering complex cellular mechanisms.
Purpose of the Study:
- To present a detailed, reproducible protocol for determining lipid-protein interactions.
- To provide a method applicable to various proteins and liposome compositions.
Main Methods:
- Preparation and characterization of liposomes using commercially available lipids.
- Formation of sucrose gradients containing liposomes and purified proteins.
- Gradient centrifugation followed by fraction collection and analysis to identify lipid-protein binding.
Main Results:
- The protocol successfully demonstrates the determination of lipid-protein interactions.
- The method is adaptable for diverse liposome formulations and protein targets.
- Liposome size is assessed to ensure protocol efficacy.
Conclusions:
- The described gradient centrifugation technique offers a robust method for studying lipid-protein interactions.
- This protocol facilitates research into the roles of lipid-protein complexes in cellular functions.
- The method's versatility supports broad applications in biochemical and cell biology research.
