Various proteomic approaches are being applied in brain tumor proteomics with regard to targeted proteins of interest, to discover phenotype specific markers which could facilitate diagnosis as well as potential antitumor drug targets. iTRAQ technology is a multiplexing protein quantitation strategy that provides relative and absolute measurements of protein abundance in complex mixtures based on the differential labeling of the proteins. Combined with membrane enrichment methodology, separation of the labeled peptides by two-dimensional liquid chromatography linked to tandem mass spectrometry and adequate data-mining, it can provide an excellent tool in search for novel isoform- and species-specific biomarkers and drug targets in various biotechnological and biomedical applications.Glioblastoma (GBM) is the most frequent primary brain tumor diagnosed in adults and remains one of the most lethal forms of human cancer. No biomarkers can distinguish different cell populations within GBMs or predict the potential of low grade gliomas to develop into malignant angiogenic gliomas. In our study, we have used iTRAQ-based technology to search for novel biomarkes of GBM.