Typical Reaction Conditions (P0732)
- Combine 1 µg of glycoprotein or 100nM oligosaccharide and H2O (if necessary) in a total reaction volume of 8 µl.
- Adding 1 µl of 10X GlycoBuffer 1 and 1 µl of 10X BSA (diluted 1:10 from 100X concentration) to make a 10 μl total reaction volume.
- Add enzyme (see below)
- Incubate reaction at 37°C for 1 hour.
- The amount of exoglycosidase enzyme required varies when different substrates are used. Start with 1-2 µl for 1 µg of glycoprotein or 100 nM of oligosacharide 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 reference the Glycobiology Double Digest Chart.
- For a more detailed characterization of several glycosidase enzymes, please reference the Detailed Characterization of Several Glycosidase Enzymes page.