title: The Use of Detergents to Purify Membrane Proteins.
abstract: Extraction of membrane proteins from biological membranes is usually accomplished with the help of detergents. This unit describes the use of detergents to solubilize and purify membrane proteins. The chemical and physical properties of the different classes of detergents typically used with biological samples are discussed. A separate section addresses the compatibility of detergents with applications downstream of the membrane protein purification process, such as optical spectroscopy, mass spectrometry, protein crystallography, biomolecular NMR, or electron microscopy. A brief summary of alternative membrane protein solubilizing and stabilizing systems is also included. Protocols in this unit include the isolation and solubilization of biological membranes and phase separation; support protocols for detergent removal, detergent exchange, and the determination of critical micelle concentration using different methods are also included.
authors.affiliation: University of Oslo, Department of Biosciences, Oslo, Norway.
authors.forename: Marcella
authors.initials: M
authors.lastname: Orwick-Rydmark
authors.affiliation: Boehringer-Ingelheim Veterinary Research Center, Hannover, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Arnold
authors.affiliation: University of Oslo, Department of Biosciences, Oslo, Norway.
authors.forename: Dirk
authors.initials: D
authors.lastname: Linke
mesh_terms: D001671:Biochemistry / Q000379:methods*; D002021:Buffers; D002845:Chromatography / Q000379:methods*; D003124:Colorimetry / Q000379:methods; D003461:Crystallography / Q000379:methods; D003902:Detergents / Q000737:chemistry*; D003956:Dialysis; D004591:Electrophoresis, Polyacrylamide Gel; D013058:Mass Spectrometry / Q000379:methods; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification*; D008823:Micelles; D008854:Microscopy, Electron / Q000379:methods; D019906:Nuclear Magnetic Resonance, Biomolecular; D017830:Octoxynol; D009994:Osmolar Concentration; D011092:Polyethylene Glycols / Q000737:chemistry; D012995:Solubility
chemical_list: D002021:Buffers; D003902:Detergents; D008565:Membrane Proteins; D008823:Micelles; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40
keywords: cloud point; detergent; detergent exchange; detergent removal; membrane protein micelle; phase separation; solubilization