Quantitative Proteomics Analysis Based on SILAC/Dimethyl Labeling
SILAC is an abbreviation for Stable Isotope Labeling by Amino Acids in Cell Culture. The SILAC experimental process involves adding light, medium, or heavy stable isotope-labeled essential amino acids lysine (Lys) and arginine (Arg) to the cell culture medium. As cells grow, newly synthesized proteins incorporate these stable isotope labels. Proteins from different groups are mixed equally, digested, and analyzed by mass spectrometry. Relative quantification is achieved by comparing the peak areas of unlabeled/labeled isotopic peaks in the MS1 spectra, while peptide sequencing in MS2 spectra allows for protein identification. Similar to SILAC, Dimethyl labeling is another isotopic protein labeling method. Dimethyl is a small molecule isotopic chemical label used to label the N-terminus or the ε-amino group of lysine in proteins or peptides after cell lysis. Both SILAC and Dimethyl labeling utilize similar mass spectrometry analysis methods, offering comparable accuracy and coverage, thus being widely used in quantitative proteomics research.
BiotechPack uses the Thermo Fisher Q Exactive HF mass spectrometry platform, Orbitrap Fusion mass spectrometry platform, and Orbitrap Fusion Lumos mass spectrometry platform combined with Nano-LC to offer a quantitative proteomics analysis service package based on SILAC/Dimethyl. You only need to inform us of your experimental objectives and send your cells to us; we will handle all subsequent project tasks, including cell culture, cell labeling, protein extraction, protein digestion, peptide separation, mass spectrometry analysis, raw data analysis, and bioinformatics analysis.
SILAC Quantitative Proteomics Experimental Process
SILAC Quantitative Proteomics Experimental Process
About Samples
1. You only need to send your frozen cells to us using dry ice, ensuring the cells can be passaged at least 5 times.
2. If you need to label the cells yourself and complete the stimulation process, you only need to send the labeled and treated cell or protein samples to us using dry ice; each sample should contain 50-100ug of protein.
Comparison of SILAC and Dimethyl Labeling Methods
Comparison of SILAC and Dimethyl Labeling Methods
Comparison of SILAC and Dimethyl Labeling Methods 2
Overall, SILAC has slightly higher reproducibility than Dimethyl because different SILAC samples are mixed before protein digestion, whereas Dimethyl labeling occurs on peptide fragments post-digestion, which is more affected by system stability. However, Dimethyl is a cost-effective labeling method that is increasingly being used.
Bilingual Project Report
In the technical report, BiotechPack provides you with a detailed bilingual report in Chinese and English, including:
1. Experimental Steps (Chinese and English)
2. Relevant Mass Spectrometry Parameters (Chinese and English)
3. Mass Spectrometry Images
4. Raw Data
5. Protein Differential Level Analysis
6. Bioinformatics Analysis
One-stop SILAC/Dimethyl Quantitative Proteomics Service
You Just Need to Order and Send Samples
BiotechPack One-stop Service Completion: Sample Processing - Instrumental Analysis - Data Analysis - Project Report
