U-Blot™ Plasma Membrane-Derived Lipid Raft Isolation Kit

Lipid rafts are small membrane domains containing a high level of cholesterol and sphingolipids. Lipid rafts have been found in the plasma membrane (PM) and internal organellar membranes such as mitochondria-associated membrane (MAMs) and endoplasmic reticulum. Lipid rafts are implicated in numerous cellular processes such as signal transduction, membrane trafficking, and protein sorting.  Lipid-modified proteins and some transmembrane proteins are concentrated in the rafts while other proteins are excluded. Lipid rafts are also found to be associated with Na⁺/K⁺ ATPase on PM. Traditional methods for lipid raft isolation involve isolation of detergent-resistant membrane subdomain from total membranous structures, which does not distinguish plasma membrane-derived and/or organelle-derived lipid rafts. Using the patented spin-column-based technologies, we have developed this kit specifically for the isolation of plasma membrane-derived lipid rafts. Larger plasma membrane vesicles are first isolated and treated with a non-ionic detergent containing buffer  followed by isolation of detergent-resistant fraction by flotation centrifugation using a tabletop microcentrifuge. Highly enriched plasma membrane-derived lipid rafts can be obtained in about 1 hour without using a traditional homogenizer and ultracentrifugation.