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Reduction and S-Carboxymethylation of Proteins: Large-Scale Method
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protocol">Protocol</div> <div class="article abstract-view" itemprop="articleBody"><span class="highwire-journal-article-marker-start"></span><h1 id="article-title-1" itemprop="headline">Reduction and S-Carboxymethylation of Proteins: Large-Scale Method</h1> <div class="contributors"> <ol class="contributor-list" id="contrib-group-1"> <li class="last" id="contrib-1"><span class="name"><a class="name-search" href="/search?author1=Richard+J.+Simpson&sortspec=date&submit=Submit">Richard J. Simpson</a></span></li> </ol> </div><span class="SOURCE">This protocol was adapted from “Peptide Mapping and Sequence Analysis of Gel-Resolved Protein,” Chapter 7, in <a href="http://www.cshlpress.com/link/proteop.htm"><em>Proteins and Proteomics</em></a> (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.</span><div class="section abstract" id="abstract-1" itemprop="description"> <h2>INTRODUCTION</h2> <p id="p-1">Intact interchain and/or intrachain disulfide linkage in a protein can present problems during proteolytic or chemical fragmentation procedures. Disulfide bonds are commonly cleaved by reducing cystine to yield cysteine residues. However, cysteine residues are highly reactive, which can complicate sequence work. In this protocol, the intact protein (>1 mg) is reduced and then S-alkylated with iodoacetic acid (or iodoacetamide). The resulting cysteine derivative S-carboxymethylcysteine (or S-carboximadomethylcysteine) is easily detectable during chemical sequencing. </p> </div><span class="highwire-journal-article-marker-end"></span></div> <div class="social-bookmarking"> <ul class="social-bookmark-links"> <li class="social-bookmarking-item social-bookmarking-item-citeulike"><a href="/external-ref?tag_url=http://cshprotocols.cshlp.org/cgi/content/long/2007/2/pdb.prot4569&title=Reduction%20and%20S-Carboxymethylation%20of%20Proteins%3A%20Large-Scale%20Method+--+Simpson%202007%20%282%29%3A%20pdb.prot4569+--+0-87969-577-3&doi=10.1101/pdb.prot4569&link_type=CITEULIKE"><img src="/shared/img/common/social-bookmarking/citeulike.gif" alt="Add to CiteULike" title="CiteULike" /></a><span class="soc-bm-link-text">CiteULike</span></li> <li class="social-bookmarking-item social-bookmarking-item-delicious"><a href="/external-ref?tag_url=http://cshprotocols.cshlp.org/cgi/content/long/2007/2/pdb.prot4569&title=Reduction%20and%20S-Carboxymethylation%20of%20Proteins%3A%20Large-Scale%20Method+--+Simpson%202007%20%282%29%3A%20pdb.prot4569+--+0-87969-577-3&doi=10.1101/pdb.prot4569&link_type=DEL_ICIO_US"><img src="/shared/img/common/social-bookmarking/delicious.gif" alt="Add to Delicious" title="Delicious" /></a><span class="soc-bm-link-text">Delicious</span></li> <li class="social-bookmarking-item social-bookmarking-item-digg"><a href="/external-ref?tag_url=http://cshprotocols.cshlp.org/cgi/content/long/2007/2/pdb.prot4569&title=Reduction%20and%20S-Carboxymethylation%20of%20Proteins%3A%20Large-Scale%20Method+--+Simpson%202007%20%282%29%3A%20pdb.prot4569+--+0-87969-577-3&doi=10.1101/pdb.prot4569&link_type=DIGG"><img src="/shared/img/common/social-bookmarking/digg.gif" alt="Add to Digg" title="Digg" /></a><span class="soc-bm-link-text">Digg</span></li> <li class="social-bookmarking-item social-bookmarking-item-facebook"><a href="/external-ref?tag_url=http://cshprotocols.cshlp.org/cgi/content/short/2007/2/pdb.prot4569&title=Reduction%20and%20S-Carboxymethylation%20of%20Proteins%3A%20Large-Scale%20Method+--+Simpson%202007%20%282%29%3A%20pdb.prot4569+--+0-87969-577-3&doi=10.1101/pdb.prot4569&link_type=FACEBOOK" class="sb-facebook" rel="external-nw"><img src="/shared/img/common/social-bookmarking/facebook.gif" alt="Add to Facebook" title="Facebook" /></a><span class="soc-bm-link-text">Facebook</span></li> <li class="social-bookmarking-item social-bookmarking-item-googleplus"><a href="/external-ref?tag_url=http://cshprotocols.cshlp.org/cgi/content/long/2007/2/pdb.prot4569&title=Reduction%20and%20S-Carboxymethylation%20of%20Proteins%3A%20Large-Scale%20Method+--+Simpson%202007%20%282%29%3A%20pdb.prot4569+--+0-87969-577-3&doi=10.1101/pdb.prot4569&link_type=GOOGLEPLUS&log_only=yes"><img src="/shared/img/common/social-bookmarking/googleplus.jpg" alt="Add to Google+" title="Google+" /></a><span class="soc-bm-link-text">Google+</span></li> <li class="social-bookmarking-item social-bookmarking-item-reddit"><a href="/external-ref?tag_url=http://cshprotocols.cshlp.org/cgi/content/long/2007/2/pdb.prot4569&title=Reduction%20and%20S-Carboxymethylation%20of%20Proteins%3A%20Large-Scale%20Method+--+Simpson%202007%20%282%29%3A%20pdb.prot4569+--+0-87969-577-3&doi=10.1101/pdb.prot4569&link_type=REDDIT"><img src="/shared/img/common/social-bookmarking/reddit.gif" alt="Add to Reddit" title="Reddit" /></a><span class="soc-bm-link-text">Reddit</span></li> <li class="social-bookmarking-item social-bookmarking-item-twitter"><a href="/external-ref?tag_url=http://cshprotocols.cshlp.org/cgi/content/long/2007/2/pdb.prot4569&title=Reduction%20and%20S-Carboxymethylation%20of%20Proteins%3A%20Large-Scale%20Method+--+Simpson%202007%20%282%29%3A%20pdb.prot4569+--+0-87969-577-3&doi=10.1101/pdb.prot4569&link_type=TWITTER"><img src="/shared/img/common/social-bookmarking/twitter.gif" alt="Add to Twitter" title="Twitter" /></a><span class="soc-bm-link-text">Twitter</span></li> </ul> <p class="social-bookmarking-help"><a href="/help/social_bookmarks.dtl">What's this?</a></p> </div><span id="related-urls"></span><div id="relmgr-related"></div> <div id="rel-related-protocol" class="relmgr-relation Related_Protocols"> <h2>Related Protocols</h2> <ul class="related-list"> <li class="cit"> <div class="cit-metadata"><span class="cit-first-element cit-section">Protocol </span><span class="cit-title">Reduction and S-Carboxymethylation of Proteins: Microscale Method </span><ul class="cit-auth-list"> <li class="first-item"><span class="cit-auth cit-auth-type-author">Richard J. 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