Accelerated Identification of Proteins by Mass Spectrometry by Employing Covalent Pre-Gel Staining with Uniblue A 英文参考文献.docVIP

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Accelerated Identification of Proteins by Mass Spectrometry by Employing Covalent Pre-Gel Staining with Uniblue A 英文参考文献.doc

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Accelerated Identification of Proteins by Mass Spectrometry by Employing Covalent Pre-Gel Staining with Uniblue A 英文参考文献

AcceleratedIdentificationofProteinsbyMass SpectrometrybyEmployingCovalentPre-GelStaining withUniblueA MarcoA.Mata-Go′mez1,MatthewT.Yasui2,ArmandoGuerrero-Rangel3,SilviaValde′s-Rodr?′guez3, RobertWinkler1,3 * 1DepartmentofBiotechnologyandFoodEngingeering,Tecnolo′gicodeMonterrey,Monterrey,NuevoLeo′n,Mexico,2SchoolofChemical,BiologicalandEnvironmental Engineering, Oregon State University, Corvallis, Oregon, United States of America, 3Department of Biotechnology and Biochemistry, CINVESTAV Unidad Irapuato, Irapuato,Guanajuato,Mexico Abstract Background:Theidentificationofproteinsbymassspectrometryisastandardmethodinbiopharmaceuticalqualitycontrol and biochemical research. Prior to identification by mass spectrometry, proteins are usually pre-separated by electrophoresis. However, current protein staining and de-staining protocols are tedious and time consuming, and thereforeprolongthesamplepreparationtimeformassspectrometry. MethodologyandPrincipalFindings:Wedevelopeda1-minutecovalentpre-gelstainingprotocolforproteins,whichdoes notrequirede-stainingbeforethemassspectrometryanalysis.Weinvestigatedtheelectrophoreticpropertiesofderivatized proteinsandpeptidesandstudiedtheirbehaviorinmassspectrometry.Further,weelucidatedthepreferredreactionof proteinswithUniblueAanddemonstratetheintegrationofthepeptidederivatizationintotypicalinformaticstools. ConclusionsandSignificance:TheUniblueAstainingmethoddrasticallyspeedsupthesamplepreparationforthemass spectrometrybasedidentificationofproteins.Theapplicationofthischemo-proteomicstrategywillbeadvantageousfor routinequalitycontrolofproteinsandfortime-criticaltasksinproteinanalysis. Citation:Mata-Go′mezMA,YasuiMT,Guerrero-RangelA,Valde′s-Rodr?′guezS,WinklerR(2012)AcceleratedIdentificationofProteinsbyMassSpectrometryby EmployingCovalentPre-GelStainingwithUniblueA.PLoSONE7(2):e31438.doi:10.1371/journal.pone.0031438 Editor:RizwanHasanKhan,AligarhMuslimUniversity,India ReceivedSeptember19,2011;AcceptedJanuary8,2012;PublishedFebruary17