title: The use of ASB-14 in combination with CHAPS is the best for solubilization of human brain proteins for two-dimensional gel electrophoresis.
abstract: Protein extraction is the most important step to reveal a proteome by Two-Dimensional Gel Electrophoresis. Usually, the urea/thiourea based standard protein extraction buffer (SB) is combined with detergents with the aim of achieving better resolution and solubilization of different classes of proteins. In order to produce better gels and achieve the greatest spot resolution of Human Brain Proteins, comparisons using 2-DE of extracted proteins from Human Brain Frontal Cortex with SB constituents (7M Urea, 2M Thiourea and 100mM DTT) were made, using different detergent compositions in the buffer. SB preparations in combination with CHAPS and ASB-14 as well as with ASB-16 (reported for the first time in 2-DE experiments) have been tested. Our results confirm that the most efficient solubilizing solution for 2-DE analysis of cytosolic and membrane Human Brain Proteins is SB combined with 4% CHAPS and 2% ASB-14.
authors.affiliation: Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), 13083-970 Campinas, SP, Brazil. daniel@proteome.ibi.unicamp.br
authors.forename: Daniel
authors.initials: D
authors.lastname: Martins-de-Souza
authors.forename: Daniel
authors.initials: D
authors.lastname: Martins
authors.forename: Bruno
authors.initials: B
authors.lastname: Menezes de Oliveira
authors.forename: Alessandro
authors.initials: A
authors.lastname: dos Santos Farias
authors.forename: Ricardo Shiniti Oka
authors.initials: RS
authors.lastname: Horiuchi
authors.forename: Cleyton
authors.initials: C
authors.lastname: Crepaldi Domingues
authors.forename: Eneida
authors.initials: E
authors.lastname: de Paula
authors.forename: Sérgio
authors.initials: S
authors.lastname: Marangoni
authors.forename: Wagner Farid
authors.initials: WF
authors.lastname: Gattaz
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Dias-Neto
authors.forename: José
authors.initials: J
authors.lastname: Camillo Novello
mesh_terms: D001622:Betaine / Q000031:analogs & derivatives* / Q000494:pharmacology; D001923:Brain Chemistry; D002793:Cholic Acids / Q000494:pharmacology*; D003902:Detergents / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D011506:Proteins / Q000737:chemistry*; D012995:Solubility
chemical_list: D002793:Cholic Acids; D003902:Detergents; D011506:Proteins; C506587:amidosulfobetaine-14; D001622:Betaine; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate