Product Description alpha-L-fucoside fucohydrolase, alpha-L-fucosidase, alpha-(1-6) fucosidase Removes non-reducing, terminal branched fucose when linked alpha-(1-6) to the core GlcNAc of a GlcNAc-GlcNAc disaccharide structure that has been labeled with a fluorescent molecule. To date, the fluorescent labels ANTS, ANDSA and 2-AA have been demonstrated to function with the fucosidase. There is no cleavage of unlabeled oligosaccharides.The one exception is that a terminal unbranched α(1-3) or α(1-4) fucose is cleaved in the absence of any reporter molecule. α(1-6) Fucosidase is useful for determining core fucosylation. Specificity α(1-6) linked core fucose when covalently attached to a reporter molecule at the reducing terminal GlcNAc. The one exception is that a terminal unbranched α(1-3) ) or α(1-4) fucose is cleaved in the absence of any reporter molecule. Reporter molecules known to support cleavage are amino-napthalene disulfonic and tri- sulfonic acids and 2-aminobenzoic acid(2AA). However, 2-aminobenzamide(2-AB)will not support cleavage. Shorter oligosaccharides such as trimannosylchitobiose are more completely digested than longer derivatives which may require longer incubation times. Source recombinant Elizabethkingia meningosepticum expressed in E. Coli
Purity Each lot of α(1-6) Fucosidase is tested for contaminating protease as follows: 10μg 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.
EC 3.2.1.51 Specific Activity >1.5 U/mg Activity >1.0 U/ml Molecular weight ~55,000 daltons
Specific Activity Assay:One unit of Fucosidase activity is defined as the amount of enzyme required to cleave 1 µmole of fucose from 4-methylumbelliferyl-alpha-L-fucoside in 1 minute at 37˚C and pH 5.0.
Contents Alpha-(1-6) fucosidase in 20 mM Tris-HCl, 25 mM NaCl,(pH 7.5). Included with 20 µL and 60 µL pack sizes: 200 µl 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 labeled 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-(1-6)-Fucosidase. 5. Incubate for 3 hours 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.
The production host strain has been extensively tested and does not produce any detectable glycosidases.
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