BlotGlyco™ Glycan Purification and Labelling Kit

• Enables the purification of glycans from biological samples (such as serum, cells and tissues) containing high levels of contaminants
• Enables the analysis of glycans within SDS-PAGE gel bands
• Purification and labelling can be completed within a single tube in approximately 5 hours

Applications

• Analysis of glycan modifications in biopharmaceuticals (research and development, quality control)
• Glycan analysis of stem cells
• Screening for biomarkers derived from biological samples
• Glycan analysis within electrophoresis bands (proteomics)

Glycans are important as biomarkers for various cellular functions and diseases; however, conventional analytical methods have made it difficult to accurately analyse complex N-linked glycans. High-purity sample preparation is essential for accurate glycan analysis using techniques such as HPLC and LC-MS.

Kit Contents

Catalog No.	Product Name	Polymer beads	Reaction spin tube	Cleanup Column	Operating Protocol
BS-45408Z	BlotGlyco 100C	100Tests	-	-	1ea
BS-45409Z	BlotGlyco 100D	100Tests	-	100	1ea
BS-45410Z	BlotGlyco 200C	200Tests	-	-	1ea
BS-45411Z	BlotGlyco 100E	100Tests	100	-	1ea
BS-45414Z	BlotGlyco 10B	10Tests	10	10	1ea
BS-45415Z	BlotGlyco 50B	50Tests	50	50	1ea

Workflow for glycan purification using BlotGyco™

By utilising the chemical bond between polymer beads—which contain a high density of Hz (hydrazide groups) for glycan capture—and the aldehyde groups at the ends of the glycans, it is possible to purify only the glycans from bio-derived samples containing a high level of contaminants, such as biological tissues and natural products, and to label them.

• Glycan chains are purified by utilising the chemical bond between the hydrazide group on the polymer beads and the aldehyde group at the terminus of the glycan chain. Consequently, glycan chains can be purified efficiently even in the presence of contaminants such as peptides.
• By fragmenting the sample proteins into peptide fragments via trypsin digestion, the efficiency of N-glycosidase can be maximised. This enables the comprehensive recovery and analysis of the glycan chains from the proteins.

Makino R, Natsuka S.: PLoS One. 2025 20(12) : e0336565. https://doi.org/10.1371/journal.pone.0336565

Optimizing the preparation of labeled N-glycans for rapid, simplified, and high-precision analysis

Background

Glycan structures show great potential as biomarkers for the early detection of diseases, as they sensitively reflect changes in cellular states. However, traditional glycan analysis has been challenging due to its complexity, time-consuming processes, and reliance on hazardous chemicals, such as those used in hydrazinolysis methods. These limitations have hindered its broader application and use in interdisciplinary research.

Research Achievements

Authors focused on developing a simple, accurate N-glycan analysis method capable of handling high-throughput sample processing. A central feature of this study is the use of pyridylamination, a fluorescent labeling technique that significantly enhances isomer separation efficiency in reversed-phase LC/MS. This approach improves the precision of glycan profiling, which is critical for identifying subtle differences in glycan structures. Through a thorough comparison of methods for N-glycan release and purification, we determined that Rapid PNGase F (New England Biolabs) and BlotGlyco™ (Sumitomo Bakelite) were the most suitable tools for this application. To further enhance accuracy, we optimized the BlotGlyco protocol by modifying the manufacturer's standard instructions, effectively reducing artifact formation during the analysis process.
Using this optimized workflow, they successfully analyzed glycan profiles from human serum, human urine, and CHO-K1 membrane fractions. This demonstrated that unique glycan structures specific to each sample type could be efficiently separated and accurately quantified. These results validate the method's capability to handle diverse biological samples while maintaining high precision and reproducibility.
This study highlights a robust and user-friendly approach to glycan analysis, which addresses key technical barriers and opens the door for broader adoption across various research fields, including biomarker discovery and glycomics. The optimized workflow offers researchers a powerful tool to explore glycan structures and their roles in biological systems, supporting advancements in diagnostics and disease understanding.

* For details, please refer to the paper

Fig. The combination of BlotGlyco™
and HPLC enables precise glycan analysis

1. Ozawa Y, et al. Time-of-Day Differences in the Plasma N‑Glycome of Normal and Obese Mice and the Effects of Dapagliflozin Administered in the Morning or at Night., Journal of Proteome Research., 25, 5(2026): 2558–2568.
   https://doi.org/10.1021/acs.jproteome.6c00081
2. Feleke DG, et al. Upregulation of Sulfated N-Glycans in Serum as Predictive Biomarkers for Early-Stage Breast Cancer., International Journal of Molecular Sciences., 26, 11 (2025):4968.
   https://doi.org/10.3390/ijms26114968
3. FURUKAWA J, et al. Comprehensive approach to structural and functional glycomics based on chemoselective glycoblotting and sequential tag conversion., Analytical chemistry., 80, 4(2008): 1094-1101.
   https://doi.org/10.1021/ac702124d
4. Shimaoka H, et al. One-pot solid-phase glycoblotting and probing by transoximization for high-throughput glycomics and glycoproteomics., Chemistry - A European Journal., 13, 6(2007) :1664-73.
   https://doi.org/10.1002/chem.200601613
5. Miura Y., et al. Rapid and Simple Solid-Phase Esterification of Sialic Acid Residues for Quantitative Glycomics by Mass Spectrometry., Chemistry - A European Journal., 13, 17(2007) :4797- 4804
   https://doi.org/10.1002/chem.200601872