  N-glycan sequencing of antibodies
 The enzymes provided are all active in a universal buffer system, GlycoBuffer 1, and can be used in

    single digests or in any combination.
  All reagents supplied are mass spectrometry compatible.

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.

Zinc  
GlycoBuffer 1  
α2-3,6,8,9 Neuraminidase A 
α2-3 Neuraminidase S  
β-N-Acetylglucosaminidase S  
β1-4 Galactosidase S  
α1-3,4,6 Galactosidase  
α1-2,4,6 Fucosidase O  
α1-2,3,6 Mannosidase

 4°C