Improved mass spectrometric proteomic profiling of the secretome of rat vascular endothelial cells. J Proteome Res 2006 Oct;5(10):2861-4
Date
10/07/2006Pubmed ID
17022658Pubmed Central ID
PMC2814535DOI
10.1021/pr060287kScopus ID
2-s2.0-33750137036 (requires institutional sign-in at Scopus site) 52 CitationsAbstract
Serum albumin contamination of cells cultured in vitro significantly impedes the mass spectrometric analysis of proteins secreted by the cells. Here we report a novel washing and culturing technique for rat vascular endothelial cells that considerably reduces the concentration of the commonly used additive for cell culture, bovine serum albumin (BSA), in the secretome of these cells. Cells are rinsed stringently and cultured for 24 h in serum-free media without appreciably impeding cell growth or viability. The percentage of BSA scans identified by tandem mass spectrometry (LC-MS/MS) in stringently rinsed cells (average 13.2%) was significantly lower than either the moderately rinsed or no rinse cell treatments (average 35.2% and 45.2% respectively). Furthermore, the stringent wash treatment allowed the confident identification of a larger portion of the secretome of rat endothelial cells by LC-MS/MS.
Author List
Pellitteri-Hahn MC, Warren MC, Didier DN, Winkler EL, Mirza SP, Greene AS, Olivier MMESH terms used to index this publication - Major topics in bold
AnimalsCattle
Cell Culture Techniques
Cells, Cultured
Culture Media
Endothelial Cells
Endothelium, Vascular
Mass Spectrometry
Proteome
Proteomics
Rats
Serum Albumin, Bovine
