N-Glycosylation Site Analysis Service

N-glycosylation is a common form of glycosylation modification in eukaryotes. Under the catalytic action of glycosyltransferases, oligosaccharide chains form amide bonds with the asparagine residues of proteins, becoming covalently attached. N-glycosylation sites refer to the amino acid residues where N-glycosylation modifications occur. The analysis of N-glycosylation sites identifies and localizes specific amino acid residues that undergo N-glycosylation in proteins through proteolysis and mass spectrometry techniques. The analysis results reveal the structure and function of proteins, which is crucial to ensuring the optimal performance of biological products. In-depth analysis of N-glycosylation sites may unveil more biological insights and guide the development of novel therapeutics.

 

MtoZ Biolabs has developed a series of reliable and high-performance glycosylation characterization methods based on liquid chromatography-high resolution mass spectrometry (LC-MS/MS). We offer comprehensive one-stop N-glycosylation site analysis services, including N-glycan analysis, N-glycosylation site identification, and detailed glycopeptide analysis workflows. Please feel free to reach out to us for further information.

 

Experimental Equipment

1. NanoLC: Vanquish NEO

2. MS: Orbitrap Fusion Lumos

 

Case Study

N-glycosylation site analysis is performed using a method that involves releasing glycans with PNGase F in heavy water and labeling with ¹⁸O. First, based on the theoretical amino acid sequence information of the protein, an appropriate protease is selected to cleave the protein into peptides. After solvent evaporation, deglycosylation is conducted in heavy water. Subsequently, LC-MS/MS is employed to identify the peptide segments containing ¹⁸O-labeled N-glycosylation sites, thereby determining the specific locations of N-glycosylation.

 

Deliverables

In the technical report, MtoZ Biolabs will provide you with detailed technical information, including:

1. Experimental Procedures

2. Relevant Mass Spectrometry Parameters

3. N-Glycosylation Site Analysis Details

4. Mass Spectrometry Images

5. Raw Data