{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,29]],"date-time":"2025-10-29T03:16:29Z","timestamp":1761707789396,"version":"build-2065373602"},"reference-count":25,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2005,4,14]],"date-time":"2005-04-14T00:00:00Z","timestamp":1113436800000},"content-version":"vor","delay-in-days":3391,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Electrophoresis"],"published-print":{"date-parts":[[1996,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>A procedure is described for structural characterization and identification of proteins, purified by either one\u2010 or two\u2010dimensional gel electrophoresis in the low picomole to femtomole range. The purified proteins are first detected in the primary gels by the sensitive reverse staining procedure described by Fernandez\u2010Patron <jats:italic>et al.<\/jats:italic> (<jats:italic>Anal. Biochem.<\/jats:italic> 1995, <jats:italic>224<\/jats:italic>, 203\u2013211) and consecutively reeluted from combined gel pieces and concentrated in the tip of a Pasteur pipette in a secondary gel matrix consisting of either sodium dodecyl sulfatepolyacrylamide or agarose. The concentrated proteins are in\u2010matrix\u2010digested and the resulting peptides are separated by reverse\u2010phase high performance liquid chromatography (HPLC) combined with microsequencing or analyzed by matrix\u2010assisted laser desorption ionization \u2014 time of flight \u2014 mass spectrometry. Protein identification is based on sequence homology or on the peptide mass pattern. The matching peptide sequences can additionally be verified by matching their measured post\u2010source decay spectra with the calculated fragmentation patterns of the isobaric candidate peptides appearing on the search list. This is done by a computer program referred to as MassFrag, described in this paper. We demonstrate that it is possible to identify protein that are only available in the femtomole range and whose sequences are stored in nonredundant protein databases or nucleotide and expressed sequence tag databases.<\/jats:p>","DOI":"10.1002\/elps.1150170513","type":"journal-article","created":{"date-parts":[[2005,5,28]],"date-time":"2005-05-28T11:16:48Z","timestamp":1117279008000},"page":"918-924","source":"Crossref","is-referenced-by-count":58,"title":["Structural analysis and identification of gel\u2010purified proteins, available in the femtomole range, using a novel computer program for peptide sequence assignment, by matrix\u2010assisted laser desorption ionization \u2014 reflectron time\u2010of\u2010fligh \u2014 mass spectrometry"],"prefix":"10.1002","volume":"17","author":[{"given":"Kris","family":"Gevaert","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jean\u2010Luc","family":"Verschelde","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Magda","family":"Puype","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jozef","family":"Van Damme","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marc","family":"Goethals","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefaan","family":"de 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