U-Blot™ Total Lipid Raft Isolation Kit for Mammalian Cells/Tissues

Lipid rafts are small membrane domains containing high cholesterol and sphingolipids. Lipid rafts are found in the plasma membrane (PM) and internal organellar membranes such as mitochondria-associated membrane (MAMs) and endoplasmic reticulum. They have been implicated in numerous cellular processes including but not limited to signal transduction, membrane trafficking, and protein sorting. Lipid-modified proteins and some transmembrane proteins are concentrated in the rafts, while others are excluded. Lipid rafts are also found to be associated with Na+/K+ ATPase on the plasma membrane. Lipid rafts are isolated traditionally by sucrose-gradient or OptiPrep gradient using ultracentrifugation, which requires a large amount of starting material and is tedious and time- consuming. This kit was developed using our proprietary spin-column-based technologies, offering a quick and easy way to isolate lipid rafts. Total membrane fraction (including PM and organelle membranes) is first isolated and treated with a non-ionic detergent-containing buffer, followed by isolation of detergent-resistant fraction by flotation centrifugation using just a tabletop microcentrifuge. Highly enriched total lipid rafts can be obtained from cultured cells/tissues in less than 90 min without using density gradient and ultracentrifugation.