MtoZ Biolabs offers the Integrated Methylation Proteomics Service that utilizes high-resolution LC-MS/MS technology to comprehensively analyze protein methylation modifications, including monomethylation, dimethylation, and trimethylation, providing life science researchers, drug development teams, and biomedical scientists with deep insights into the role of protein methylation in key biological processes such as signal transduction, gene regulation, and cellular function while supporting the advancement of disease mechanism research and drug development.

The Principle of Protein Methylation Analysis

Protein methylation is a post-translational modification (PTM) in which a methyl group (-CH₃) is covalently added to specific amino acid residues in a protein, most commonly lysine (Kme) and arginine (Rme). This modification does not alter the amino acid sequence but can affect protein activity, interactions, and signal regulation. Types of methylation include monomethylation, dimethylation, and trimethylation, with each modification corresponding to different biological functions.

Biggar, KK. et al. Nat Rev Mol Cell Biol. 2015.

Figure 1. Lys and Arg methylation

Mass spectrometry (MS) detection of protein methylation relies on mass shift identification, modification fragment evidence reading, and residue-level localization. The classic mass increment for methyl groups is +14.0156 Da (monomethylation), +28.0313 Da (dimethylation), and +42.0469 Da (trimethylation). These precise measurable m/z shifts are direct indicators of the modification types. Commonly monitored residues include lysine and arginine, where lysine methylation does not alter charge but affects peptide hydrophobicity and fragmentation, while arginine dimethylation can be differentiated by fragment ion patterns to distinguish between symmetric and asymmetric states.

During data acquisition, high-resolution LC-MS/MS platforms (such as Orbitrap, TOF, or FTICR) are used to detect precursor and product ions, ensuring reliable modification mass shifts, co-eluted peptide separation at the same retention time, and database search-based site localization. In MS/MS fragmentation data, methylated peptides exhibit b/y fragment ion series that provide evidence for modification localization, with neutral loss trends, peak intensity distributions, and retention time behavior helping to confirm the logical stability of modification states, enabling highly reliable identification and quantification of protein methylation.

Integrated Methylation Proteomics Service at MtoZ Biolabs

The Integrated Methylation Proteomics Service is a comprehensive methylation analysis solution provided by MtoZ Biolabs, supporting both target protein-level and complex proteome-level analysis. We offer complete identification of protein methylation modifications, including monomethylation, dimethylation, and trimethylation, with a focus on key amino acid residues such as lysine and arginine. Specific services include:

• Methylation-Modified Protein or Peptide Identification: Using high-resolution LC-MS/MS technology, we accurately identify methylation sites in proteins, including lysine methylation (Kme) and arginine methylation (Rme), and detect different methylation states (e.g., Kme1, Kme2, Kme3, Rme1, Rme2).
• Relative Quantification of Methylated Peptides: Based on LC-MS/MS data, we provide relative quantification analysis of methylated peptides, comparing methylation levels across different experimental conditions or sample groups.
• Absolute Quantification of Methylated Peptides: Using stable isotope labeling techniques (such as SILAC or TMT), we perform absolute quantification analysis of methylated peptides, helping to accurately assess methylation level differences and monitor subtle changes.

Through these analyses, MtoZ Biolabs' service helps researchers gain deeper insights into the role and regulatory mechanisms of protein methylation in various biological processes, especially in signal transduction, gene expression regulation, and protein interactions.

Workflow of Integrated Methylation Proteomics Service

Why Choose MtoZ Biolabs

• High-resolution mass spectrometry platforms (Orbitrap, Q Exactive, and Fusion Lumos systems)
• Mature sample enrichment system
• Strong compatibility across sample types
• One-time fee, transparent pricing

Sample Submission Guidelines

For more information, please refer to Sample Submission Guidelines for Proteomics and Sample Submission Guidelines for Metabolomics.

Deliverables

• Raw data files from LC-MS/MS
• Methylation site identification and quantification results
• Summary report
• High-resolution images and chromatograms
• Data structure and format support

FAQs

Q1: What types of samples are suitable?

MtoZ Biolabs accepts cell lysates, tissue extracts, purified proteins, recombinant/fusion proteins, and research-grade biofluids, ensuring the samples are clear, with low detergent and low background interference.

Q2: What is the service's general workflow?

Q3: What data formats are provided?

MtoZ Biolabs provides results in multiple standard formats to ensure compatibility with various analysis and visualization tools. Deliverables typically include:

•  Raw data files from LC-MS/MS (e.g., .RAW files)
•  Processed data tables (CSV or Excel format) containing identified and quantified methylated peptides
•  Summary reports in PDF format, including analytical methods, quality control results, and key findings
•  Figures and spectra in high-resolution image formats (TIFF or PNG)

Additional file formats or data structures can be provided upon request to meet specific research or publication requirements.

Q4: How should I prepare my samples?

To ensure accurate and reproducible results for Integrated Methylation Proteomics, MtoZ Biolabs recommends preparing your samples according to the following guidelines:

• Sample Type: We accept a variety of biological samples, including cell lysates, tissue extracts, serum, plasma, and purified proteins or peptides.
• Sample Purity: Samples should be free from particulates and low in salts, detergents, or organic solvents that could interfere with chromatographic separation or ionization during MS detection.
• Volume Requirement: Please provide at least 500 μL of liquid sample or 100 mg of solid material. Additional volume may be required for replicate analyses.
• Storage and Shipping: Store biological samples at −80°C. Ship samples on dry ice in leak-proof containers to maintain sample integrity.
• Documentation: Include detailed information about sample origin, preparation method, buffer composition, and the desired analysis type.

If you're unsure about your sample preparation, our technical team is available for personalized pre-analysis consultation and can provide a sample submission guide to ensure optimal preparation and shipping conditions.

Start Your Project with MtoZ Biolabs

Contact MtoZ Biolabs today to begin your project and let our professional team provide precise and reliable methylation analysis services for your research.