Product Description 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.
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.
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.
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
EC 3.2.1.51 Specific Activity >2 U/mg Activity 0.5 U/ml Molecular weight 40,000 daltons
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.
Contents 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)
Formulation The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, 25 mM NaCl pH 7.5.
Suggested usage 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.
Storage Store enzyme at 4˚C. Stability Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce 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. |