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 prote...

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Published in:Briefings in functional genomics & proteomics 2007-03, Vol.6 (1), p.70-75
Main Authors: Martins, Daniel, Menezes de Oliveira, Bruno, dos Santos Farias, Alessandro, Oka Horiuchi, Ricardo Shiniti, Crepaldi Domingues, Cleyton, de Paula, Eneida, Marangoni, Sérgio, Farid Gattaz, Wagner, Dias-Neto, Emmanuel, Novello, José Camillo
Format: Article
Language:English
Subjects:
Betaine - analogs & derivatives
Betaine - pharmacology
Brain
Brain Chemistry
Cholic Acids - pharmacology
Detergents - pharmacology
Electrophoresis, Gel, Two-Dimensional - methods
Humans
Proteins
Proteins - chemistry
Proteomics
Scientific method
Solubility
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Summary: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.
ISSN:1473-9550
2041-2649
1477-4062
2041-2657
DOI:10.1093/bfgp/elm009