Mitochondrial SILAC & Dimethyl Analysis

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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.

Service Flow

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.

Technical characteristics

• The labeling effect of the living cell horizontal labeling technique is stable, and its labeling efficiency is not affected by the lysate. The marking efficiency is high.
• The demand for samples is small, usually tens of micrograms of the protein.
• Mass spectrometry can be used to identify and quantify many kinds of proteins at the same time.
• Multiple samples are mixed, separated, digested, and identified at the same time, the subsequent experiments had the same effect on the samples, reduced the influence of experimental operation and equipment, and had precision and reproducibility.
• The labeling uses in vivo labeling technology, which is close to the real state of the sample.

Our Advantages

• Rich experience in mitochondrial quantitative proteomics research
• Outstanding research team
• Unique integrated service for mitochondrial research
• Reliable data and results
• Rapid turnaround and cost-effective

Creative Biogene has the most professional team to serve customers around the world. If you have any questions about the content of this service, please feel free to contact us. We look forward to your contact.

Reference

1. Wang X., et al. “ SILAC–based quantitative MS approach for real-time recording protein-mediated cell-cell interactions.”Sci Rep, 2018, 8: 8441.
2. Boersema P., et al. “ Multiplex peptide stable isotope dimethyl labeling for quantitative proteomics.” Nature Protocols, 2009, 4: 484-494.

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