Typical Reaction Conditions for α1-3, 4 Fucosidase (P0769)
- Combine 1 μg of glycoprotein or 100 nM of oligosaccharide and H2O (if necessary) to make an 8 μl total reaction volume.
- 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 1 μl of α1-3,4 Fucosidase.
- 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.
- Higher enzyme concentrations and longer incubation times may be required for the removal of α1-3 and α1-4 fucose residues from complex N-glycans.
- α1-3,4 Fucosidase can cleave the α1-3 fucose on the outer arm of N-linked glycans but is unable to remove the core α1-3 fucose. erial, let the reaction go overnight.