title: An efficient solubilization buffer for plant proteins focused in immobilized pH gradients.
abstract: The solubilization of a large array of proteins before electrophoresis itself is a very critical point for proteomic analyses. We compared the efficiency of several different solubilization buffers. From this work, we defined a very efficient solubilization buffer, including two chaotropes, two reducing agents (R2), two detergents (D2), and two kinds of carrier ampholytes in combination. This so-called R2D2 buffer (5 M urea, 2 M thiourea, 2% 3-[(3-cholamidopropyl) dimethyl-ammonio]-1-propane-sulfonate, 2% N-decyl-N,N-dimethyl-3-ammonio-1-propane-sulfonate, 20 mM dithiothreitol, 5 mM Tris(2-carboxyethyl) phosphine, 0.5% carrier ampholytes 4-6.5, 0.25% carrier ampholytes 3-10) proved to be very efficient for a large range of different samples and allowed us to obtain two-dimensional gels of high resolution and quality.
authors.affiliation: INRA / INA-PG / UPS / CNRS UMR8120, Station de Génétique Végétale, Ferme du Moulon, 91190 Gif-sur-Yvette, France. mechin@moulon.inra.fr
authors.forename: Valérie
authors.initials: V
authors.lastname: Méchin
authors.forename: Luciano
authors.initials: L
authors.lastname: Consoli
authors.forename: Martine
authors.initials: M
authors.lastname: Le Guilloux
authors.forename: Catherine
authors.initials: C
authors.lastname: Damerval
mesh_terms: D002021:Buffers; D003902:Detergents / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods; D010940:Plant Proteins / Q000737:chemistry*; D011506:Proteins / Q000032:analysis*; D018892:Proton-Motive Force; D019163:Reducing Agents / Q000494:pharmacology; D016622:Silver Staining
chemical_list: D002021:Buffers; D003902:Detergents; D010940:Plant Proteins; D011506:Proteins; D019163:Reducing Agents