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Saudi Journal of Medical and Pharmaceutical Sciences (SJMPS)
Volume-3 | Issue-12 | 1308-1312
Original Research Article
Identification of Cancer Marker Vinculin from Caco-2 Cancer Cell By Using UPLC/MS Method
Engin Koçak, Kevser Biberoğlu, Sacide Altınöz
Published : Dec. 30, 2017
DOI : 10.36348/sjmps.2017.v03i12.007
Abstract
Vinculin is one the most common proteins that have been studied commonly in recent years. Vinculin has anti-tumor effect and loss of vinculin causes development of many cancer types. Because of this Vinculin is a marker and it shoulde be evaluated for invitro and clinical studies. LC/MS based proteomics is the main tool to analyze hundreds of proteins in very short time because it offers high sensitivity and accuracy. Today nano LC is the min tool for LC/MS proteomics researches but UPLC/MS based systems have been developed and offered more robust and easy way for proteomics. In this study we used normal flow UPLC/Q-TOF method to analyze caco-2 colon cancer cell line. One of our targets for this study was to identify Vinculin as a cancer marker. We digested proteins from caco-2 cells and analyzed with UPLC/MS. Recorded MS/MS data was used with Maxquant bioinformatic system. We identified 5 unique peptides that belonged to Vinculin. We used 20 ppm mass tolerance for amino acid sequence matching with uniprot human database. We made three raplicate to observe identification reprocubility. Also We used label free quantification method and calculated vinculin intensity
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