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2013 | 1 | 28-37

Article title

A sample preparation method for micro-scale membrane
proteome analysis


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Detergents are widely used to improve the solubilization and extraction of
hydrophobic membrane proteins in proteomics. Since most detergents
are not compatible with subsequent steps of analysis, the removal of
detergents from samples, especially those in micro-scale amounts,
is a worthy topic of investigation. In this paper, we present a novel
polyvinylidene difluoride (PVDF) membrane-mediated sample preparation
method for micro-scale membrane proteome analysis, using a rat liver
cell membrane-enriched fraction as model material. The proteins in the
fraction were extracted in a 2 % sodium dodecyl sulfate (SDS) solution and
the protein solution was applied on a piece of PVDF membrane followed
by drying and repeated washing in order to remove SDS and other salts.
Quantitative determination indicated that about 84% of the SDS in the
sample was removed and protein loss was less than 10%. Four methods
were used and compared for digesting the proteins adsorbed on PVDF
membrane. Dimethyl formamide (DMF)-assisted digestion was the most
effective with regard to the identification of membrane proteins, particularly
the highly hydrophobic multi-transmembrane proteins. These results
demonstrate that PVDF membrane-aided sample cleanup combined with
DMF-assisted digestion has potential utility in the micro-scale membrane
proteome analysis.







Physical description


09 - 2 - 2013
22 - 11 - 2013
24 - 08 - 2013


  • Key Laboratory of Protein Chemistry
    and Developmental Biology of National
    Education Committee, College of Life
    Sciences, Hunan Normal University,
    Changsha, 410081, P. R. China
  • Key Laboratory of Protein Chemistry
    and Developmental Biology of National
    Education Committee, College of Life
    Sciences, Hunan Normal University,
    Changsha, 410081, P. R. China
  • Key Laboratory of Protein Chemistry
    and Developmental Biology of National
    Education Committee, College of Life
    Sciences, Hunan Normal University,
    Changsha, 410081, P. R. China
  • Key Laboratory of Protein Chemistry
    and Developmental Biology of National
    Education Committee, College of Life
    Sciences, Hunan Normal University,
    Changsha, 410081, P. R. China


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