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It relies on the following steps: (i) gel electrophoresis separation of proteins, (ii) fast in\u2010gel protein digestion with trypsin, (iii) <jats:sup>18<\/jats:sup>O\u2010labeling through the decoupled method, (iv) quantification through selected peptides previously chosen using the <jats:sup>18<\/jats:sup>O inverse labeling approach and that, finally, (v) it takes advantage of software specifically developed to select the peptides that will drive the quantification of the protein in an automated mode. We have accurately quantified the following six proteins: glycogen phosphorylase, BSA, ovalbumin, carbonic anhydrase, trypsin inhibitor, and \u03b1\u2010lactalbumin. As a case study we have quantified the protein vitellogenin in plasma of <jats:italic>Cyprinus carpio<\/jats:italic> exposed to high levels of estrogens. The proposed new protocol was validated against the traditional ELISA method; both were found to provide comparable results (non\u2010parametric Mann\u2013Whitney <jats:italic>U<\/jats:italic>\u2010test).<\/jats:p>","DOI":"10.1002\/elps.201000251","type":"journal-article","created":{"date-parts":[[2010,9,29]],"date-time":"2010-09-29T06:11:25Z","timestamp":1285740685000},"page":"3407-3419","source":"Crossref","is-referenced-by-count":10,"title":["A novel <sup>18<\/sup>O inverse labeling\u2010based workflow for accurate bottom\u2010up mass spectrometry quantification of proteins separated by gel electrophoresis"],"prefix":"10.1002","volume":"31","author":[{"given":"Hugo M.","family":"Santos","sequence":"first","affiliation":[]},{"given":"Daniel","family":"Glez\u2010Pe\u00f1a","sequence":"additional","affiliation":[]},{"given":"Miguel","family":"Reboiro\u2010Jato","sequence":"additional","affiliation":[]},{"given":"Florentino","family":"Fdez\u2010Riverola","sequence":"additional","affiliation":[]},{"given":"M\u00e1rio S.","family":"Diniz","sequence":"additional","affiliation":[]},{"given":"Carlos","family":"Lodeiro","sequence":"additional","affiliation":[]},{"given":"Jos\u00e9\u2010Luis","family":"Capelo\u2010Mart\u00ednez","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2010,9,29]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/45350"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1522-2683(19990101)20:4\/5<664::AID-ELPS664>3.0.CO;2-A"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1124619"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/msb.2008.61"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr800836c"},{"key":"e_1_2_6_7_2","doi-asserted-by":"publisher","DOI":"10.1007\/s12035-009-8071-2"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1002\/prca.200780128"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio736"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.200300885"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac802484d"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr900528j"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.M900026-MCP200"},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.talanta.2009.04.053"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr8009879"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr800467r"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.trac.2006.05.015"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1322"},{"key":"e_1_2_6_19_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac035286f"},{"key":"e_1_2_6_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1044-0305(03)00207-1"},{"key":"e_1_2_6_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2010.03.003"},{"key":"e_1_2_6_22_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr050405o"},{"key":"e_1_2_6_23_2","doi-asserted-by":"publisher","DOI":"10.1002\/rcm.1290091216"},{"key":"e_1_2_6_24_2","doi-asserted-by":"publisher","DOI":"10.1093\/bfgp\/eln041"},{"key":"e_1_2_6_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac990298f"},{"key":"e_1_2_6_26_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac010043d"},{"key":"e_1_2_6_27_2","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.200900197"},{"key":"e_1_2_6_28_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac001404c"},{"key":"e_1_2_6_29_2","doi-asserted-by":"publisher","DOI":"10.1023\/A:1008986522208"},{"key":"e_1_2_6_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ecoenv.2004.11.004"},{"key":"e_1_2_6_31_2","volume":"10","author":"Glez\u2010Pena D.","year":"2009","journal-title":"BMC Bioinformatics"},{"key":"e_1_2_6_32_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr8009098"},{"key":"e_1_2_6_33_2","doi-asserted-by":"publisher","DOI":"10.1002\/rcm.3214"},{"key":"e_1_2_6_34_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr700850w"},{"key":"e_1_2_6_35_2","doi-asserted-by":"publisher","DOI":"10.1021\/pr025572s"},{"key":"e_1_2_6_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.talanta.2008.03.013"},{"key":"e_1_2_6_37_2","doi-asserted-by":"publisher","DOI":"10.1002\/rcm.525"},{"key":"e_1_2_6_38_2","first-page":"74","volume":"11","author":"Speicher K.","year":"2000","journal-title":"J. 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