The use of ASB-14 in combination with CHAPS is the best for solubilization of human brain proteins for two-dimensional gel electrophoresis

  • Brief Funct Genomic Proteomic. 2007 Mar;6(1):70-5. doi: 10.1093/bfgp/elm009.
Daniel Martins-de-Souza  1 ,  Bruno Menezes de Oliveira ,  Alessandro dos Santos Farias ,  Ricardo Shiniti Oka Horiuchi ,  Cleyton Crepaldi Domingues ,  Eneida de Paula ,  Sérgio Marangoni ,  Wagner Farid Gattaz ,  Emmanuel Dias-Neto ,  José Camillo Novello
Affiliations
  • 1. Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), 13083-970 Campinas, SP, Brazil. [email protected]
PMID: 17556486 DOI: 10.1093/bfgp/elm009
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.