The use of ASB-14 in combination with CHAPS is the best for solubilization of human brain proteins for two-dimensional gel electrophoresis.
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.
Martins-de-Souza
Daniel
D
Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), 13083-970 Campinas, SP, Brazil. daniel@proteome.ibi.unicamp.br
Martins
Daniel
D
Menezes de Oliveira
Bruno
B
dos Santos Farias
Alessandro
A
Horiuchi
Ricardo Shiniti Oka
RS
Crepaldi Domingues
Cleyton
C
de Paula
Eneida
E
Marangoni
Sérgio
S
Gattaz
Wagner Farid
WF
Dias-Neto
Emmanuel
E
Camillo Novello
José
J
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
D002793:Cholic Acids; D003902:Detergents; D011506:Proteins; C506587:amidosulfobetaine-14; D001622:Betaine; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate