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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Comparison of Protein-Extraction Methods for Gills of the Shore Crab, <italic>Carcinus maenas</italic>
(L.), and Application to 2DE</title>
<author><name sortKey="Panchout, Francois" sort="Panchout, Francois" uniqKey="Panchout F" first="François" last="Panchout">François Panchout</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Letendre, Julie" sort="Letendre, Julie" uniqKey="Letendre J" first="Julie" last="Letendre">Julie Letendre</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Bultelle, Florence" sort="Bultelle, Florence" uniqKey="Bultelle F" first="Florence" last="Bultelle">Florence Bultelle</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Denier, Xavier" sort="Denier, Xavier" uniqKey="Denier X" first="Xavier" last="Denier">Xavier Denier</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Rocher, Beatrice" sort="Rocher, Beatrice" uniqKey="Rocher B" first="Béatrice" last="Rocher">Béatrice Rocher</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Chan, Philippe" sort="Chan, Philippe" uniqKey="Chan P" first="Philippe" last="Chan">Philippe Chan</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff3">Plate-forme de Protéomique Pissaro, Institute for Research and Innovation in Biomedicine</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Vaudry, David" sort="Vaudry, David" uniqKey="Vaudry D" first="David" last="Vaudry">David Vaudry</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff3">Plate-forme de Protéomique Pissaro, Institute for Research and Innovation in Biomedicine</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff4">Neuronal and Neuroendocrine Differentiation and Communication, DC2N, Inserm U982, University of Rouen, 76821 Mont-Saint-Aignan, France</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Durand, Fabrice" sort="Durand, Fabrice" uniqKey="Durand F" first="Fabrice" last="Durand">Fabrice Durand</name>
<affiliation><nlm:aff wicri:cut="; and" id="aff2">Faculty of Sciences and Technologies, University of Le Havre, 76058 Le Havre cedex, France</nlm:aff>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">24294114</idno>
<idno type="pmc">3792702</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3792702</idno>
<idno type="RBID">PMC:3792702</idno>
<idno type="doi">10.7171/jbt.13-2404-002</idno>
<date when="2013">2013</date>
<idno type="wicri:Area/Pmc/Corpus">000186</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Comparison of Protein-Extraction Methods for Gills of the Shore Crab, <italic>Carcinus maenas</italic>
(L.), and Application to 2DE</title>
<author><name sortKey="Panchout, Francois" sort="Panchout, Francois" uniqKey="Panchout F" first="François" last="Panchout">François Panchout</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Letendre, Julie" sort="Letendre, Julie" uniqKey="Letendre J" first="Julie" last="Letendre">Julie Letendre</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Bultelle, Florence" sort="Bultelle, Florence" uniqKey="Bultelle F" first="Florence" last="Bultelle">Florence Bultelle</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Denier, Xavier" sort="Denier, Xavier" uniqKey="Denier X" first="Xavier" last="Denier">Xavier Denier</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Rocher, Beatrice" sort="Rocher, Beatrice" uniqKey="Rocher B" first="Béatrice" last="Rocher">Béatrice Rocher</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Chan, Philippe" sort="Chan, Philippe" uniqKey="Chan P" first="Philippe" last="Chan">Philippe Chan</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff3">Plate-forme de Protéomique Pissaro, Institute for Research and Innovation in Biomedicine</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Vaudry, David" sort="Vaudry, David" uniqKey="Vaudry D" first="David" last="Vaudry">David Vaudry</name>
<affiliation><nlm:aff wicri:cut=", and" id="aff3">Plate-forme de Protéomique Pissaro, Institute for Research and Innovation in Biomedicine</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff4">Neuronal and Neuroendocrine Differentiation and Communication, DC2N, Inserm U982, University of Rouen, 76821 Mont-Saint-Aignan, France</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Durand, Fabrice" sort="Durand, Fabrice" uniqKey="Durand F" first="Fabrice" last="Durand">Fabrice Durand</name>
<affiliation><nlm:aff wicri:cut="; and" id="aff2">Faculty of Sciences and Technologies, University of Le Havre, 76058 Le Havre cedex, France</nlm:aff>
</affiliation>
</author>
</analytic>
<series><title level="j">Journal of Biomolecular Techniques : JBT</title>
<idno type="ISSN">1524-0215</idno>
<idno type="eISSN">1943-4731</idno>
<imprint><date when="2013">2013</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<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>
</front>
</TEI>
<pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Biomol Tech</journal-id>
<journal-id journal-id-type="iso-abbrev">J Biomol Tech</journal-id>
<journal-id journal-id-type="pmc">jbt</journal-id>
<journal-id journal-id-type="publisher-id">jbt</journal-id>
<journal-title-group><journal-title>Journal of Biomolecular Techniques : JBT</journal-title>
</journal-title-group>
<issn pub-type="ppub">1524-0215</issn>
<issn pub-type="epub">1943-4731</issn>
<publisher><publisher-name>Association of Biomolecular Resource Facilities</publisher-name>
<publisher-loc>Bethesda, MD</publisher-loc>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">24294114</article-id>
<article-id pub-id-type="pmc">3792702</article-id>
<article-id pub-id-type="publisher-id">13-2404-002</article-id>
<article-id pub-id-type="doi">10.7171/jbt.13-2404-002</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Comparison of Protein-Extraction Methods for Gills of the Shore Crab, <italic>Carcinus maenas</italic>
(L.), and Application to 2DE</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Panchout</surname>
<given-names>François</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
<xref ref-type="corresp" rid="cor1"></xref>
</contrib>
<contrib contrib-type="author"><name><surname>Letendre</surname>
<given-names>Julie</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Bultelle</surname>
<given-names>Florence</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Denier</surname>
<given-names>Xavier</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Rocher</surname>
<given-names>Béatrice</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chan</surname>
<given-names>Philippe</given-names>
</name>
<xref ref-type="aff" rid="aff3"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Vaudry</surname>
<given-names>David</given-names>
</name>
<xref ref-type="aff" rid="aff3"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff4"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Durand</surname>
<given-names>Fabrice</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>4</sup>
</xref>
</contrib>
<aff id="aff1"><sup>1</sup>
Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222, and</aff>
<aff id="aff2"><sup>4</sup>
Faculty of Sciences and Technologies, University of Le Havre, 76058 Le Havre cedex, France; and</aff>
<aff id="aff3"><sup>2</sup>
Plate-forme de Protéomique Pissaro, Institute for Research and Innovation in Biomedicine, and</aff>
<aff id="aff4"><sup>3</sup>
Neuronal and Neuroendocrine Differentiation and Communication, DC2N, Inserm U982, University of Rouen, 76821 Mont-Saint-Aignan, France</aff>
</contrib-group>
<author-notes><corresp id="cor1">A<sc>ddress correspondence to</sc>
: François Panchout, <addr-line>Laboratory of Ecotoxicology–Aquatic Environments (LEMA, EA 3222), University of Le Havre, B.P. 540, 76058 Le Havre Cedex, France</addr-line>
(Phone: <phone>+33(0)2 32 85 99 09</phone>
; Fax: <fax>+33(0)2 32 74 45 05</fax>
; E-mail: <email>francois.panchout@univ-lehavre.fr</email>
).</corresp>
</author-notes>
<pub-date pub-type="ppub"><month>12</month>
<year>2013</year>
</pub-date>
<pub-date pub-type="pmc-release"><month>12</month>
<year>2013</year>
</pub-date>
<pmc-comment> PMC Release delay is 0 months and 0 days and was based on the
. </pmc-comment>
<volume>24</volume>
<issue>4</issue>
<fpage>218</fpage>
<lpage>223</lpage>
<permissions><copyright-statement>© 2013 Association of Biomolecular Resource Facilities</copyright-statement>
<copyright-year>2013</copyright-year>
</permissions>
<self-uri xlink:title="pdf" xlink:type="simple" xlink:href="jbt00413000218.pdf"></self-uri>
<abstract><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>
</abstract>
<kwd-group><kwd>2D gel electrophoresis</kwd>
<kwd>hard tissue</kwd>
<kwd>chitin</kwd>
<kwd>crustacean</kwd>
</kwd-group>
</article-meta>
</front>
</pmc>
</record>
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