By a News Reporter-Staff News Editor at Life Science Weekly -- A new study on Proteomics is now available. According to news reporting from Oslo, Norway, by NewsRx journalists, research stated, "The proteomics field has shifted over recent years from two-dimensional gel electrophoresis (2-DE)-based approaches to SDS-PAGE or gel-free workflows because of the tremendous developments in isotopic labeling techniques, nano-liquid chromatography, and high-resolution mass spectrometry. However, 2-DE still offers the highest resolution in protein separation."
The news correspondents obtained a quote from the research from the University of Oslo, "Therefore, we combined stable isotope labeling with amino acids in cell culture of controls and apoptotic HeLa cells with 2-DE and the subsequent analysis of tryptic peptides via nano-liquid chromatography coupled to an LTQ-Orbitrap mass spectrometer to obtain quantitative data using the methods with the highest resolving power on all levels of the proteomics workflow. More than 1,200 proteins with more than 2,700 protein species were identified and quantified from 816 Coomassie Brilliant Blue G-250 stained 2-DE spots. About half of the proteins were identified and quantified only in single 2-DE spots. The majority of spots revealed one to five proteins; however, in one 2-DE spot, up to 23 proteins were identified. Only half of the 2-DE spots represented a dominant protein with more than 90% of the whole protein amount. Consequently, quantification based on staining intensities in 2-DE gels would in approximately half of the spots be imprecise, and minor components could not be quantified. These problems are circumvented by quantification using stable isotope labeling with amino acids in cell culture. Despite challenges, as shown in detail for lamin A/C and vimentin, the quantitative changes of protein species can be detected."
According to the news reporters, the research concluded: "The combination of 2-DE with high-resolution nano-liquid chromatography-mass spectrometry allowed us to identify proteomic changes in apoptotic cells that would be unobservable using any of the other previously employed proteomic workflows."
For more information on this research see: High resolution quantitative proteomics of HeLa cells protein species using stable isotope labeling with amino acids in cell culture(SILAC), two-dimensional gel electrophoresis(2DE) and nano-liquid chromatograpohy coupled to an LTQ-OrbitrapMass spectrome. Molecular and Cellular Proteomics, 2013;12(2):529-38. (American Society for Biochemistry and Molecular Biology - www.asbmb.org; Molecular and Cellular Proteomics - www.mcponline.org/)
Our news journalists report that additional information may be obtained by contacting B. Thiede, The Biotechnology Centre of Oslo, University of Oslo, Gaustadalleen 21, 0349 Oslo, Norway. Additional authors for this research include C.J. Koehler, M. Strozynski, A. Treumann, R. Stein, U. Zimny-Arndt, M. Schmid and P.R Jungblut (see also Proteomics).
Keywords for this news article include: Oslo, Norway, Europe, Peptides, Proteins, Hela Cells, Proteomics, Amino Acids.
Our reports deliver fact-based news of research and discoveries from around the world. Copyright 2014, NewsRx LLC