title: Use of universal stable isotope labeling by amino acids in cell culture (SILAC)-based selected reaction monitoring (SRM) approach for verification of breast cancer-related protein markers.
abstract: Mass spectrometry-based proteomics facilitates high-throughput discovery of protein markers for diagnosis and treatment of breast cancer patients. Hundreds of putative prognostic and predictive markers are being identified every year, but only a very small proportion of them can be validated as clinically relevant markers. A quantitative and cost-efficient verification method is highly desirable to pick up real "nuggets" from the "sand." To fulfill these criteria, we previously introduced a stable isotope labeling by amino acids in cell culture (SILAC)-based selected reaction monitoring (SRM) approach for studying breast cancer-related protein markers. Here we describe a hands-on protocol of using this SILAC-SRM method for verification of breast cancer-related markers, which can also be used for verification of protein markers in other types of solid tumor tissues.
authors.affiliation: Department of Medical Oncology, Laboratory of Breast Cancer Genomics and Proteomics, Erasmus University Medical Center, Erasmus MC Cancer Institute, Wytemaweg 80, Be-401, 2040, 3000, CA, Rotterdam, The Netherlands.
authors.forename: Ning Qing
authors.initials: NQ
authors.lastname: Liu
authors.forename: Lennard J M
authors.initials: LJ
authors.lastname: Dekker
authors.forename: Martijn M
authors.initials: MM
authors.lastname: Van Duijn
authors.forename: Arzu
authors.initials: A
authors.lastname: Umar
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D014408:Biomarkers, Tumor / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D005260:Female; D006801:Humans; D007553:Isotope Labeling; D009363:Neoplasm Proteins / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D000596:Amino Acids; D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins