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

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1156; p. 307
• Main Authors: Liu, Ning Qing, Dekker, Lennard J M, Van Duijn, Martijn M, Umar, Arzu
• Format: Journal Article
• Language: English
• Published: United States 2014
• Subjects: Amino Acids - chemistry; Biomarkers, Tumor - metabolism; Breast Neoplasms - metabolism; Breast Neoplasms - pathology; Cell Line, Tumor; Chromatography, Liquid; Female; Humans; Isotope Labeling; Neoplasm Proteins - metabolism; Tandem Mass Spectrometry
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-0685-7_21

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.