Typical Reaction Conditions for α1-3, 4, 6 Galactosidase (P0747)

Optimal incubation times and enzyme concentrations must be determined empirically for a particular substrate. Typical reaction conditions are as follows:
  1. Combine 1 μg of glycoprotein or 100 nM of oligosaccharide and H2O (if necessary) to make an 8 μl total reaction volume.

  2. Add 1 μl of 10X GlycoBuffer 1 and 1 μl of 10X rHSA (diluted 1:10 from 100X concentration) to make a 10 μl total reaction volume.

  3. Add 1 μl of α1-3,4,6 Galactosidase.

  4. Incubate at 37°C for 1 hour. 

  • Reactions may be scaled-up linearly to accommodate larger reaction volumes.
  • The amount of exoglycosidase enzyme required varies when different substrates are used. Start with 1-2 μl for 1ug of glycoprotein or 100 nM of oligosaccharide for one hour in a 10-25 μl reaction. If there is still undigested material, let the reaction go overnight.