Alpha (1-3,4) Fucosidase cleaves branched non-reducing terminal fucose, linked α(1-3) or α(1-4) to the N-acetylglucosamine of terminal Gal-GlcNAc disaccharide structures.
Part Number – Amount of Enzyme
E-F134 – 60 µLs¹
E-F134-20 – 20 µLs¹
E-F134-200 – 200 µls²
¹ includes buffer
² includes enzyme only
alpha-L-fucoside fucohydrolase, alpha-L-fucosidase, alpha-(1-3,4) fucosidase
Alpha (1-3,4) Fucosidase cleaves branched non-reducing terminal fucose, linked α(1-3) or α(1-4) to the N-acetylglucosamine of terminal Gal-GlcNAc disaccharide structures. The presence of sialic acid (but not fucose) linked to the galactose will block cleavage.
For removing core fucose linked α-(1-6) to the core GlcNAc of a GlcNAc-GlcNAc disaccharide structure we recommend our Alpha-(1-6) Fucosidase (E-F006).
α(1-3, 4) Fucosidase is useful for:
nbsp;nbsp;Fucose linkage determination
nbsp;nbsp;Deglycosylating glycoproteins with Lewis structures
Source Isolated from Xanthamonas manihotis
Alpha-(1-3,4)-Fucosidase in 20 mM Tris-HCl, 25 mM NaCl,(pH 7.5).
Included with 20 µL and 60 µL pack sizes:
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
Formulation The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, 25 mM NaCl pH 7.5.
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 Assay 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.
Purity Each lot of α(1-3, 4) Fucosidase is tested for contaminating activities by incubating
the enzyme for 24 hours at 37°C with the appropriate substrates; the detection limit of this assay is 5 μU/mL (IUB). A passing lot will have no detectable activity.
For the protease assay, 10 μg of denatured BSA is incubated for 24 hours with 2 μL of enzyme. Analysis of the BSA band after SDS-PAGE should show no evidence of degradation.
Stability Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.