O-GlcNAcylation Analysis Service
MtoZ Biolabs provides O-GlcNAcylation Analysis Service to deliver accurate identification, mapping, and quantification of O-linked N-acetylglucosamine modifications on proteins. Leveraging Orbitrap Fusion Lumos and Q Exactive HF mass spectrometers, combined with optimized enrichment antibodies and specialized bioinformatics pipelines, this service helps researchers resolve one of the most dynamic and regulatory protein modifications in cell biology. By providing reliable, publication-ready datasets, MtoZ Biolabs empowers scientists to investigate the role of O-GlcNAcylation in cellular signaling, metabolic regulation, and disease pathogenesis.
What is O-GlcNAcylation?
O-GlcNAcylation is a reversible post-translational modification in which a single N-acetylglucosamine (GlcNAc) residue is attached to serine or threonine residues of nuclear, cytoplasmic, and mitochondrial proteins. Unlike classical glycosylation that takes place in the endoplasmic reticulum and Golgi, O-GlcNAcylation occurs exclusively inside cells and acts as a metabolic and stress-sensitive "molecular switch." This modification influences enzyme activity, protein stability, transcriptional activity, subcellular localization, and protein–protein interactions. It is widely conserved across eukaryotes and has been observed in certain bacteria, fungi, and viruses. Dysregulation of O-GlcNAcylation is implicated in cancer, diabetes, cardiovascular disease, and neurodegenerative disorders, making it an important focus of modern biomedical research.
Services at MtoZ Biolabs
1. Targeted Protein O-GlcNAcylation Analysis
MtoZ Biolabs offers targeted analysis of O-GlcNAcylation on client-specified proteins. Clients may submit one or multiple targets for site-level detection of O-GlcNAc modifications. Following proteolytic digestion, O-GlcNAc-modified peptides are enriched using optimized workflows and analyzed by high-resolution LC-MS/MS to accurately identify and quantify modification sites on the designated proteins.
2. O-GlcNAcylation Proteomics
For comprehensive profiling, MtoZ Biolabs provides O-GlcNAcylation proteomics to assess O-GlcNAc modifications across the entire proteome. Through large-scale LC-MS/MS combined with specialized enrichment of O-GlcNAcylated peptides, this service enables systematic mapping and quantitative comparison of modification patterns in complex biological samples, supporting global or condition-dependent studies.
Analysis Workflow
The O-GlcNAcylation Analysis Service at MtoZ Biolabs follows a validated workflow:
1. Sample Preparation: Protein extraction under conditions that preserve labile O-GlcNAc modifications.
2. Enzymatic Digestion: Proteins are digested with proteases to generate peptides suitable for downstream enrichment.
3. Enrichment: High-specificity immunoprecipitation or lectin-based approaches are applied to selectively isolate O-GlcNAcylated peptides.
4. LC-MS/MS Acquisition: Orbitrap-based mass spectrometry ensures high-resolution, sensitive detection and site localization.
5. Data Analysis: Bioinformatics pipelines annotate O-GlcNAcylation sites, quantify modification levels, and integrate functional insights.
6. Quality Control: Rigorous QC metrics, including TIC evaluation and replicate consistency, ensure reproducibility.
Hu, W. et al. RSCAdv. 2022.
Figure 1. Quantitative Approaches for O-GlcNAcylation Analysis
Why Choose MtoZ Biolabs?
✔ Advanced Instrumentation: Orbitrap Fusion Lumos and Q Exactive HF ensure high-resolution and accurate detection of O-GlcNAc sites.
✔ Enrichment Expertise: Antibody- and lectin-based workflows improve recovery of low-abundance O-GlcNAcylated peptides.
✔ Comprehensive Analysis: From site-specific mapping to bioinformatics interpretation, offering complete end-to-end solutions.
✔ Collaborative Support: Scientific experts provide consultation to interpret results and guide follow-up research.
✔ One-Time-Charge: Our pricing is transparent, no hidden fees or additional costs.
Sample Submission Suggestions
To ensure optimal analysis, MtoZ Biolabs recommends:
Sample Types: Accepted materials include animal tissues, plant tissues, cultured cells, and purified proteins.
Amounts: Standard submission is 200–300 mg tissue, 6–8 × 107 cells, or 10–20 mg purified protein.
Preservation: Store samples at -80°C and ship on dry ice.
Preparation Notes: Avoid detergents, glycerol, or salts that may interfere with LC-MS/MS analysis.
MtoZ Biolabs provides a detailed sample submission guide and consultation to support researchers during preparation and shipping.
Applications
Our O-GlcNAcylation Analysis Service supports multiple research applications:
Metabolic Regulation: Study O-GlcNAc as a nutrient sensor linking cellular energy state to protein function.
Neurobiology: Explore O-GlcNAcylation in neuronal plasticity and its relevance to Alzheimer’s and Parkinson’s disease.
Cancer Research: Investigate O-GlcNAc-driven changes in transcription, cell cycle control, and oncogenic signaling.
Cardiovascular Studies: Examine O-GlcNAcylation in stress adaptation and cardiac signaling pathways.
PTM Crosstalk: Evaluate interplay between O-GlcNAcylation and phosphorylation to reveal regulatory complexity.
FAQ
Q1: What strategies are used to improve the enrichment efficiency of O-GlcNAcylated peptides?
Enrichment is a critical step due to the low abundance of O-GlcNAcylation compared with other modifications. At MtoZ Biolabs, we use antibody-based immunoprecipitation combined with complementary lectin enrichment methods to maximize specificity and recovery. This dual approach ensures a more comprehensive capture of O-GlcNAc-modified peptides, improving downstream MS detection and site localization.
Q2: How do you address the labile nature of O-GlcNAc modifications during sample preparation and analysis?
O-GlcNAc residues are chemically labile and prone to loss during conventional workflows. To preserve modification integrity, we optimize protein extraction buffers, apply gentle digestion protocols, and employ fragmentation methods such as electron transfer dissociation (ETD) in LC-MS/MS. These strategies minimize modification loss and enhance the confidence of site assignments.
O-GlcNAcylation is a highly dynamic and reversible modification that plays essential roles in signaling, metabolism, and disease. Its complexity and low abundance make advanced analytical strategies necessary. MtoZ Biolabs provides a comprehensive O-GlcNAcylation Analysis Service that integrates enrichment expertise, cutting-edge mass spectrometry, and bioinformatics support. By delivering accurate, reproducible, and biologically meaningful results, we help researchers across academia and industry unlock the regulatory functions of O-GlcNAcylation and drive innovation in health and disease research.
What Could be Included in the Report?
1. Comprehensive Experimental Details
2. Materials, Instruments, and Methods
3. Total Ion Chromatogram & Quality Control Assessment
4. Data Analysis, Preprocessing, and Estimation
5. Bioinformatics Analysis
6. Raw Data Files