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Shotgun Mass Spectrometry Workflow Combining IEF and LC-MALDI-TOF/TOF

We present a high throughput shotgun mass spectrometry workflow using a bidimensional peptide fractionation procedure consisting of isoelectric focusing and RP-HPLC prior to mass spectrometric analysis, with the aim of optimizing peptide separation and protein identification. As part of the workflow...

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Detalles Bibliográficos
Autores principales: Maccarrone, Giuseppina, Turck, Christoph W., Martins-de-Souza, Daniel
Formato: Texto
Lenguaje:English
Publicado: Springer US 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2836250/
https://www.ncbi.nlm.nih.gov/pubmed/20091422
http://dx.doi.org/10.1007/s10930-009-9227-7
Descripción
Sumario:We present a high throughput shotgun mass spectrometry workflow using a bidimensional peptide fractionation procedure consisting of isoelectric focusing and RP-HPLC prior to mass spectrometric analysis, with the aim of optimizing peptide separation and protein identification. As part of the workflow we used the ‘Isotope-Coded Protein Labeling’ (ICPL) method for accurate relative quantitation of protein expression. Such workflow was successfully applied to a comparative proteome analysis of schizophrenia versus healthy control brain tissues and can be an alternative to proteome researches.