title: Optimal concentrations of N-decanoyl-N-methylglucamine and sodium dodecyl sulfate allow the extraction and analysis of membrane proteins.
abstract: We studied the extraction and analysis of integral membrane proteins possessing hydrophobic and hydrophilic domains and found that a nonionic detergent called MEGA-10, used in lysis buffers, had a superior extraction effect compared to most conventional detergents. A sodium dodecyl sulfate (SDS) concentration of >0.4% (w/v) in the sample buffer was crucial for those proteins to be clearly analyzed by electrophoresis and Western blotting. Furthermore, MEGA-10 had the tendency to maximally extract proteins around its critical micelle concentration (CMC) of 0.24% (w/v). These solutions can greatly assist functional investigations of membrane proteins in the proteomics era.
authors.affiliation: Department of Life Science, National Taiwan Normal University, Taipei 116, Taiwan.
authors.forename: Jen-Hua
authors.initials: JH
authors.lastname: Chuang
authors.forename: Yu-Jing
authors.initials: YJ
authors.lastname: Kao
authors.forename: Neil B
authors.initials: NB
authors.lastname: Ruderman
authors.forename: Li-Chu
authors.initials: LC
authors.lastname: Tung
authors.forename: Yenshou
authors.initials: Y
authors.lastname: Lin
mesh_terms: D015153:Blotting, Western / Q000379:methods; D002021:Buffers; D004586:Electrophoresis / Q000379:methods; D005227:Fatty Acids / Q000737:chemistry*; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D008823:Micelles; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D012995:Solubility; D013501:Surface-Active Agents / Q000737:chemistry*
chemical_list: D002021:Buffers; D005227:Fatty Acids; D008565:Membrane Proteins; D008823:Micelles; D013501:Surface-Active Agents; D012967:Sodium Dodecyl Sulfate; C042221:decanoyl-N-methylglucamide; D005944:Glucosamine