Stable isotope labeling by amino acids in cell culture (SILAC) technique uses medium containing light, medium, or heavy essential amino acids (mainly Lys and Arg) to culture cells. Through the normal metabolism of cells, the newly synthesized proteins are labeled with stable isotopes. The same amount of protein samples of different treatments are mixed, and the quantitative and qualitative results of the proteins could be obtained by SDS-PAGE separation, gel cutting, and LC-MS/ MS analysis after enzyme digestion.
Figure 1. Quantitative analysis of Triple-SILAC and Spike-in SILAC. ( Wang X., et al, 2018)
Similar to SILAC, Dimethyl labeling is also a method of isotope protein labeling. Dimethyl is a small molecular isotope chemical marker, which marks the N-terminal of the lytic protein or peptide or the ε-amino group of lysine by chemical labeling so that the protein/ peptide is labeled with different Dimethyl tags. In addition to the differences in labeling methods, the mass spectrometry analysis methods of the two SILAC/Dimethyl markers are the same and have similar analytical accuracy and breadth. [ Learn more about iTRAQ ]
Figure 2. Comparison of dimethyl labeling with iTRAQ and SILAC. ( Boersema P., et al, 2009)
Creative Biogene uses Thermo Fisher's Q ExactiveHF mass spectrometry platform, Orbitrap Fusion mass spectrometry platform, Orbitrap Fusion Lumos mass spectrometry platform combined with Nano-LC to provide the label-free quantitative proteome analysis service. Tell us the goal of your experiment and send us your cells, and we will be responsible for all the follow-up matters of the project. Including cell culture, cell labeling, protein extraction, protease digestion, peptide separation, mass spectrometry analysis, mass spectrometry original data analysis, bioinformatics analysis.
Creative Biogene will provide you with a detailed technical report when the project is delivered, including experimental steps, relevant MS parameters, details of identified amino acids, MS images (high resolution), and detailed raw data. Creative Biogene always provides you with customized services for mitochondrial research, and our one-stop research and analysis platform can meet all your research needs.
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