Application data
- Breast cancer is the most common form of cancer in Asian women. In Singapore, breast cancer is increasing at an average rate of 3% per annum.
- In this study, we aim to identify the potential tumor biomarkers by comparing the protein expression profiles among different breast cancer cell lines (MCF-7, HCC-38) and control cell lines (CRL-2072).
- A large-scale and high-throughput proteomic platform was employed for the project. About 1,000 protein spots were visualized by the narrow pH range gels (pH 4-7 and 6-9) for each cell line and a total of 249 differentially displayed spots were selected and identified by MS analysis using the Xcise as part of the gel processing.
- Peptide mass fingerprinting was achieved using the curved field reflectron instrument AXIMA-CFR plus™ (Shimadzu Biotech). Peptide sequencing was performed using a quadrupole ion trap mass spectrometer AXIMA-QIT™ (Shimadzu Biotech). Proteins were identified using the Mascot database searching engine (Matrix Science).
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Xcise was used to perform gel acquisition imaging, protein spot cutting and digestion, peptide extraction and deposition onto the MALDI target plate. To each gel plug, 0.1µg trypsin (Promega Corporation) was used for digestion at 30 ºC overnight with shaking. After cleaning and concentrating with m-C18 ZipTip (Millipore), the tryptic-peptide mixture was mixed with matrices (CHCA:DHB = 1:1, 10 mg/ml) and loaded onto a 384-well MALDI sample plate (Shimadzu Biotech).
Presented at HUPO 2004 - Bejing
Systematic Proteome Analysis of Breast Cancer Cell Lines
K.L. Ou1,2, S.Y. Soon1,2, C.C. Yau1, D. Kesuma1, A. Tan1, J. Liu1, W. Chen1, P. Tan1 and H. Jikuya1,2
1 Agenica Research Pte Ltd, Singapore.
2 Shimadzu (Asia Pacific) Pte Ltd, Singapore.
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