Typical Reaction Conditions for α2-3 Neuraminidase (P0728)

1. Combine 1 μg of glycoprotein or 100 nM of oligosaccharide and H₂O (if necessary) in a total reaction volume of 8 μl.
2. Add 1 μl of GlycoBuffer 1 (10X) and 1 μl of BSA (10X) (diluted 1:10 from 100X concentration) to make a 10 μl total reaction volume.
3. Add enzyme (see below)
4. Incubate at 37°C for 1 hour.

Notes

• The amount of exoglycosidase enzyme required varies when different substrates are used. Start with 1-2 μl for 1 ug of glycoprotein or 100 nM of oligosachride for one hour in a 10-25 μl reaction. If there is still undigested material, let the reaction go overnight.
• The reaction can be scaled up linearly
• Several exo- and endo-glycosidases can be used together in one digest. For buffer recommendations, please referance the Glycobiology Double Digest Chart .
• For a more detailed characterization of several glycosidase enzymes, please reference the Detailed Characterization of Several Glycosidase Enzymes page.