Part number E-F134
60 µl aliquot of enzyme (30 mU) in 20 mM Tris-HCl, 25 mM NaCl,(pH 7.5).
200 µl 5x Reaction Buffer 5.0 (250 mM sodium phosphate, pH 5.0)
Specific Activity >2 U/mg
Activity 0.5 U/ml
Molecular weight 40,000 daltons
1. Add up to 1 nmole of oligosaccharide to a tube.
2. Add de-ionized water to a total of 15 µl.
3. Add 4 µl of 5x Reaction Buffer 5.0.
4. Add 1 µl of Alpha-(1-3,4)-Fucosidase.
5. Incubate for 1 hour at 37˚C.
Specifictity Non-reducing terminal branched fucose when linked alpha-(1-3) or alpha-(1-4) to GlcNAc of a Gal-GlcNAc disaccharide structure. The presence of sialic acid (but not fucose) linked to the galactose will block cleavage.
Specific Activity One unit of Fucosidase activity is defined as the amount of enzyme required to cleave 1 µmole of fucose from Lewis X trisaccharide, 4-methylumbelliferyl glycoside in 1 minute at 37˚C and pH 5.0. Lewis X trisaccharide is Gal Beta-(1-4)[Fuc alpha-(1-3)]GlcNAc.
Storage Store enzyme at 4˚C.