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

Daniel MartinsdeSouza | Zendy; B. Menezes de Oliveira | Zendy; A. dos Santos Farias | Zendy; Ricardo Shiniti Oka Horiuchi | Zendy; Cleyton C. Domingues | Zendy; Eneida de Paula | Zendy; Sérgio Marangoni | Zendy; Wagner F. Gattaz | Zendy; Emmanuel DiasNeto | Zendy; J. C. Novello | Zendy

Open Access

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

Author(s) -

Daniel MartinsdeSouza,

B. Menezes de Oliveira,

A. dos Santos Farias,

Ricardo Shiniti Oka Horiuchi,

Cleyton C. Domingues,

Eneida de Paula,

Sérgio Marangoni,

Wagner F. Gattaz,

Emmanuel DiasNeto,

J. C. Novello

Publication year - 2007

Publication title -

briefings in functional genomics and proteomics

Language(s) - English

Resource type - Journals

eISSN - 1477-4062

pISSN - 1473-9550

DOI - 10.1093/bfgp/elm009

Subject(s) - chaps , thiourea , chromatography , urea , extraction (chemistry) , electrophoresis , human brain , biology , proteome , membrane protein , gel electrophoresis , solubilization , biochemistry , chemistry , membrane , organic chemistry , neuroscience

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.

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