The N-Glycan Sequencing Kit consists of seven well characterized, highly pure, recombinant exoglycosidase enzymes selected to simplify the process of characterizing typical N-linked glycan structures.
The N-Glycan Sequencing Kit consists of seven well characterized, highly pure, recombinant exoglycosidase enzymes selected to simplify the process of characterizing typical N-linked glycan structures. The extreme diversity of glycan structures on a glycoprotein makes elucidation of the profile challenging and often a number of orthogonal approaches are employed to verify the individual structures. The use of sequential or tandem exoglycosidase digestion of oligosaccharides followed by mass spectrometry or capillary electrophoresis analysis provides detailed carbohydrate sequence information and resolves ambiguities. The exoglycosidases are all active in a universal buffer system, GlycoBuffer 1, and can be used in single digests or in combination.
This kit is compatible with N-linked glycans released from a variety of CHO and murine derived antibodies, as well as N-linked glycans released from other glycoproteins. Optimal incubation times and enzyme concentration may vary for more complex glycans. The recommended enzyme quantity is sufficient for digestion of up to 130 pmol of released glycans (equivalent to released glycans from approximately 10 µgs of antibody) labeled with 2AA, 2AB or procainamide. Other commercially available “Instant labels” may require a larger quantity of enzyme or a longer incubation time for complete digestion.
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|GlycoBuffer 1||-20||10 X|
|α2-3,6,8,9 Neuraminidase A||-20||20,000 units/ml|
|α2-3 Neuraminidase S||-20||8,000 units/ml|
|β-N-Acetylglucosaminidase S||4||4,000 units/ml|
|β1-4 Galactosidase S||-20||8,000 units/ml|
|α1-3,4,6 Galactosidase||4||8,000 units/ml|
|α1-2,4,6 Fucosidase O||4||2,000 units/ml|
|α1-2,3,6 Mannosidase||4||2,000 units/ml|
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