Abstract
Proteins associate with membranes in a number of ways. Is the membrane an integral membrane protein? A peripheral protein? Does it have a glycosylphosphoinositol (GPI) tail? Is it associated with caveolae-derived plasma membrane domains? This unit describes methods for determining how a protein of interest associates with the membrane including extraction with alkaline carbonate, urea, high salt, or Triton X-114. GPI linkage is probed using PI-PLC cleavage. There is a method for detergent solubilization of Triton-X 100 insoluble integral and GPI-linked proteins. Caveolae membranes can be isolated using Triton X-100 or a detergent-free method.
MeSH terms
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Animals
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Caveolae / chemistry
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Cell Fractionation / methods
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Cell Membrane / chemistry*
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Cell Membrane / drug effects
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Detergents / pharmacology
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Glycosylphosphatidylinositols / chemistry
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Humans
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Hydrogen-Ion Concentration
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Membrane Proteins / chemistry
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Membrane Proteins / isolation & purification
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Octoxynol
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Osmolar Concentration
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Phosphoinositide Phospholipase C
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Polyethylene Glycols
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Potassium Chloride
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Protease Inhibitors
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Protein Binding
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Proteins / chemistry*
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Proteins / isolation & purification
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Sodium Chloride
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Urea
Substances
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Detergents
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Glycosylphosphatidylinositols
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Membrane Proteins
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Protease Inhibitors
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Proteins
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Polyethylene Glycols
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Sodium Chloride
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Potassium Chloride
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Urea
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Octoxynol
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Nonidet P-40
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Phosphoinositide Phospholipase C