Typical Reaction Conditions for α1-2,3 Mannosidase (P0729)
- Combine 1 μg of glycoprotein or 100 nM of oligosaccharide and H2O (if necessary) in a total reaction volume of 8 μl.
- Add 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 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 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 referance the Detailed Characterization of Several Glycosidase Enzymes page.