Comparison of Protein-Extraction Methods for Gills of the Shore Crab, Carcinus maenas (L.), and Application to 2DE
Identifieur interne : 000443 ( France/Analysis ); précédent : 000442; suivant : 000444Comparison of Protein-Extraction Methods for Gills of the Shore Crab, Carcinus maenas (L.), and Application to 2DE
Auteurs : François Panchout ; Julie Letendre ; Florence Bultelle ; Xavier Denier ; Béatrice Rocher ; Philippe Chan ; David Vaudry [France] ; Fabrice Durand [France]Source :
- Journal of Biomolecular Techniques : JBT [ 1524-0215 ] ; 2013.
Abstract
As it is well-established that protein extraction constitutes a crucial step for two-dimensional electrophoresis (2DE), this work was done as a prerequisite to further the study of alterations in the proteome in gills of the shore crab
Url:
DOI: 10.7171/jbt.13-2404-002
PubMed: 24294114
PubMed Central: 3792702
Affiliations:
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PMC:3792702Le document en format XML
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<front><div type="abstract" xml:lang="en"><p>As it is well-established that protein extraction constitutes a crucial step for two-dimensional electrophoresis (2DE), this work was done as a prerequisite to further the study of alterations in the proteome in gills of the shore crab <italic>Carcinus maenas</italic>
under contrasted environmental conditions. Because of the presence of a chitin layer, shore crab gills have an unusual structure. Consequently, they are considered as a hard tissue and represent a challenge for optimal protein extraction. In this study, we compared three published extraction procedures for subsequent applications to 2DE: the first one uses homogenization process, the second one included an additional TCA-acetone precipitation step, and finally, the third one associated grinding in liquid nitrogen (N<sub>2</sub>
) and TCA-acetone precipitation. Extracted proteins were then resolved using 1DE and 2DE. Although interesting patterns were obtained using 1DE with the three methods, only the one involving grinding in liquid N<sub>2</sub>
and TCA-acetone precipitation led to proper resolution after 2DE, showing a good level of reproducibility at technical (85%) and biological (84%) levels. This last method is therefore proposed for analysis of gill proteomes in the shore crab.</p>
</div>
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<name sortKey="Denier, Xavier" sort="Denier, Xavier" uniqKey="Denier X" first="Xavier" last="Denier">Xavier Denier</name>
<name sortKey="Letendre, Julie" sort="Letendre, Julie" uniqKey="Letendre J" first="Julie" last="Letendre">Julie Letendre</name>
<name sortKey="Panchout, Francois" sort="Panchout, Francois" uniqKey="Panchout F" first="François" last="Panchout">François Panchout</name>
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