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 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.
Maccarrone
Giuseppina
G
Max Planck Institute of Psychiatry, Proteomics and Biomarkers, Kraepelinstrasse 2, 80804, Munich, Germany.
Turck
Christoph W
CW
Martins-de-Souza
Daniel
D
D001923:Brain Chemistry; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D007553:Isotope Labeling; D013058:Mass Spectrometry / Q000379:methods*; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D012559:Schizophrenia / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D010455:Peptides