Lipid particles found in circulating extracellular fluids such as blood or lymph are essential for cellular homeostasis, metabolism and survival. Such particles provide essential lipids and fats which enable cells to synthesize new membranes and regulate different biochemical pathways. Imbalance in lipid particle metabolism can cause pathological states such as atherosclerosis. Here, elevated low-density lipoprotein (LDL) accumulation leads to fat-filled lesions or plaques in arterial walls. In this chapter, we provide a detailed set of protocols for the rapid and safe purification of lipid particles from human blood using high-speed ultracentrifugation. We provide a detailed set of assays for further analysis of the biochemical and cellular properties of these lipid particles. By combining these assays, we can better understand the complex roles of different lipid particles in normal physiology and disease pathology.
Keywords: Gradient purification; HDL; Human embryonic kidney 293T (HEK293T) cells; Human umbilical vein endothelial cells (HUVECs); LDL; Lipid particles; SDS-PAGE; VLDL.
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.