| Description: | Lipid rafts are small membrane domains containing high level of cholesterol and sphingolipids. Lipid rafts have been found in 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 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 table top microcentrifuge. Highly enriched plasma membrane derived lipid rafts can be obtained in about 1 hour without using traditional homogenizer and ultracentrifugation.Lipid rafts are small membrane domains containing high level of cholesterol and sphingolipids. Lipid rafts have been found in 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 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 table top microcentrifuge. Highly enriched plasma membrane derived lipid rafts can be obtained in about 1 hour without using traditional homogenizer and ultracentrifugation.Lipid rafts are small membrane domains containing high level of cholesterol and sphingolipids. Lipid rafts have been found in 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 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 table top microcentrifuge. Highly enriched plasma membrane derived lipid rafts can be obtained in about 1 hour without using traditional homogenizer and ultracentrifugation. |
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