Submit Your Request Now

N-Glycan Linkage Analysis by HILIC-UHPLC

Usually it is not necessary to determine the glycosidic linkages because they rarely cause effect on the function of glycoproteins. However, due to the high complexity of glycan structures of glycoprotein, assessing the configuration and position of glycosidic linkages of the N-glycan is helpful. Up to now, there are five common N-glycan linkages known. Among these five linkages, N-acetylglucosamine to asparagine (GlcNAcβ1-Asn) is the most common one. Other types of linkages to Asn include: glucose in laminin of both mammals and Archaea, N-acetylgalactosamine (GalNAc) in Archaea, rhamnosein bacteria. And a report shows that glucose linkage to argninein a sweet corn.

N-glycans are released from glycoproteins by PNGase F reaction. The released N-glycans are labeled with 2-aminobenzamide (2-AB), followed by digestion with multiple exoglycosidases. Finally, glycans are analyzed by the Hydrophilic Interaction Chromatography (HILIC) with a fluorescence and mass spectrometric detection. Several steps are involved in N-Glycan Linkage analysis:

Sample preparation
N-Glycans release
2-aminobenzamide (2-AB) labeling and the labeled glycans are reacted sequentially with a series of exoglycosidases.
UPLC-HILIC-FLD based analysis;
Additionally, confirmation the detailed glycan structures by MS/MS fragmentation can be done if required.

*What does Endoglycosidase used for?
Endoglycosidase releases oligosaccharides from glycoproteins or glycolipids. Normally it does not release oligosaccharides from conjugated protein and lipid molecules. Endoglycosidase functions by breaking the glycosidic bonds between two sugar monomer in the polymer.
Endoglycosidase H: release oligo mannose and hybrid N-glycans
Endoglycosidase F: release simple biantennary N-glycans

How to place an order:
How To Place your Order

*If your organization requires signing of a confidentiality agreement, please contact us by email

As one of the leading companies in providing omics analysis service, Creative Proteomics provides reliable glycomics analysis service for our customers. Contact us for more detailed information!

Proteomics Sample Submission Guidelines

Ensure your samples are prepared and submitted correctly by downloading our comprehensive Proteomics Sample Submission Guidelines. This document provides detailed instructions and essential information to facilitate a smooth submission process. Click the link below to access the PDF and ensure your submission meets all necessary criteria.

Download

* For Research Use Only. Not for use in diagnostic procedures.

Our customer service representatives are available 24 hours a day, 7 days a week. Inquiry

From Our Clients

"I recently used their proteomics service for a project analyzing protein interactions in yeast models. The team was very responsive and helped clarify the methodology they employed, which made me feel confident in the results. The data quality was solid, with clear identification of several key proteins involved in our study. Their thorough analysis enabled me to pinpoint specific interactions that I hadn't considered before, which significantly improved the direction of my research. I appreciate their professionalism and support throughout the process."

Sarah Thompson, University of California, Berkeley

"Our lab collaborated with them on a project studying cancer biomarkers. The proteomics analysis provided was detailed and focused, specifically highlighting the differential expression of proteins between healthy and tumor samples. Their clear explanations of the data helped my team understand the biological implications. I also appreciated their willingness to revise the reports based on our feedback, ensuring that we had everything we needed for our publication. This collaborative spirit was invaluable."

Emily Rodriguez, Stanford University

"Our lab worked with them on a project studying the effects of diet on gut microbiota using proteomics. They used a label-free quantification method to analyze proteins in fecal samples before and after dietary intervention. The results showed significant changes in protein expression linked to microbial activity. This was pivotal for our hypothesis about diet-microbiota interactions. The clarity of their data presentation made it easy for our team to integrate these findings into our ongoing research."

Dr. Lisa Wong, University of Toronto

"My experience with Creative Proteomics during the mass spectrometry analysis was excellent. We sent in human saliva and mouse brain tissue samples, which they expertly analyzed using both LC-MS and GC-MS techniques. The results were invaluable, revealing key metabolites in the saliva and identifying biomarkers linked to brain function in the brain tissue."

Dr. Emily Carter, Senior Research Scientist

"The overall service from Creative Proteomics was outstanding. They made the entire process seamless and efficient, allowing us to focus on our research. We worked with leaf and root samples from various Arabidopsis genotypes for targeted metabolomics analysis. Their thorough profiling of primary and secondary metabolites gave us important insights into how the plants respond metabolically to environmental stress."

Dr. Laura Henderson, Plant Physiologist

"We had a pleasant collaboration with Creative Proteomics on mass spectrometry analysis of lipids. They conducted a detailed analysis of lipid species, providing us with important insights into lipid metabolism and its relationship with metabolic syndrome disease states."

Dr. Sarah Mitchell, Research Scientist

Online Inquiry

Please submit a detailed description of your project. We will provide you with a customized project plan to meet your research requests. You can also send emails directly to for inquiries.

Great Minds Choose Creative Proteomics