Alpha-(1-3,4) Fucosidase

Cat. No.: A490000

GRADE & PURITY Specific Activity ＞2 U/mg;Activity 0.5 U/ml

Overview

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.

α(1-3, 4) Fucosidase is useful for:

nbsp;nbsp;Fucose linkage determination

nbsp;nbsp;Deglycosylating glycoproteins with Lewis structures

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)

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.

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.

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.

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.

Specifications

Product Name

Alpha-(1-3,4) Fucosidase

Synonyms

α-L-Fucoside fucohydralase

Specifications & Purity

Specific Activity ＞2 U/mg;Activity 0.5 U/ml

Enzyme Commission Number

3.2.1.51

Molecule Type

Enzyme

Storage and Shipping

Concentration

Specific Activity ＞2 U/mg;Activity 0.5 U/ml

Storage

Store at 2-8°C

Shipped In

Wet ice