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α1-2,3,4,6 Fucosidase

Catalog # Concentration Size List Price Quantity Your Price
P0748L 8000 units/ml 2000 units
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P0748S 8000 units/ml 400 units
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Catalog # Size List Price Your Price
P0748L 2000 units
P0748S 400 units
Please Inquire
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

α1-2,3,4,6 Fucosidase is a broad specificity exoglycosidase that catalyzes the hydrolysis of terminal, non-reducing α1-2, α1-3, α1-4 and α1-6 linked fucose residues from oligosaccharides.

  • Recombinant enzyme with no detectable endoglycosidase or other exoglycosidase contaminating activities
  • Glycerol -free for optimal performance in HPLC and mass spectrometry analysis
  • ≥95% purity, as determined by SDS-PAGE and intact ESI-MS
  • Optimal activity and stability for up to 12 months
α1-2,3,4,6 Fucosidase, cloned from bovine kidney, is a broad specificity exoglycosidase that catalyzes the hydrolysis of α1-2, α1-3, α1-4 and α1-6 linked fucose residues from oligosaccharides. α1-2,3,4,6 Fucosidase cleaves α1-2 and α1-6 fucose residues more efficiently than other linkages, and has slight activity towards α1-3 fucose residues.
Product Source
Cloned from bovine kidney and expressed in E. coli (1).

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave > 95% of the α-L-fucose from 1 nmol of Fuc α1-2Galα1-4Glc- 7-amino-4-methyl-coumarin (AMC), in 1 hour at 37°C in a total reaction volume of 10 μl.

Unit Definition Assay
Two fold dilutions of α1-2,3,4,6 Fucosidase are incubated with 1 nmol AMC-labeled substrate in 1X GlycoBuffer 1 supplemented with 100 μg/ml BSA in a 10 μl reaction. The reaction mix is incubated at 37°C for 1 hour. Separation of reaction products are visualized via thin layer chromatography (2).

Reaction Conditions

1X GlycoBuffer 1
Supplement with Purified BSA
Incubate at 37°C

1X GlycoBuffer 1
5 mM CaCl2
50 mM sodium acetate
(pH 5.5 @ 25°C)

Storage Buffer

20 mM Tris-HCl
50 mM NaCl
1 mM EDTA
pH 7.5 @ 25°C

Heat Inactivation

100°C for 10 minutes

Molecular Weight

Theoretical: 51800 daltons

Specific Activity

72,000 units/mg

Companion Products
Notes
  • 8 units of enzyme wereinactivated by incubation at 100°C for 10 minutes.
References
  • Vainauskas, S., New England Biolabs, Inc. Unpublished observation
  • Wong-Madden, S.T. and Landry, D. (1995). Glycobiology 5. 19-28.
Web Tools
FAQs
Tech Tips
    • Repeated freeze/thaw cycles may reduce activity. Recommended storage temperature is 4°C
    • Using 1-2 µl is a good starting point for a 1 hr incubation of 1µg of glycoprotein or 100 nM of oligosaccharide.
    • Longer incubation times (4 hours to overnight) may be needed for complex, branched oligosaccharide substrates.
    • α1-2,3,4,6 Fucosidase will cleave branched α1-4 and α1-6 fucose residues but will not cleave branched α1-2 fucose residues.
    • α1-2,3,4,6 Fucosidase will partially cleave linear and branched α1-3 Fucose residues with low efficiency.
    • The storage buffer does not contain BSA; however, we recommend supplementing the reaction with BSA for optimal enzyme activity
Citations
Publications
  • Albrecht, S., Vainauskas, S., Stöckmann, H., McManus, C., Taron, C.H. and Rudd, P.M. (2016). Comprehensive Profiling of Glycosphingolipid Glycans Using a Novel Broad Specificity Endoglycoceramidase in a High-Throughput Workflow. Anal Chem. May 3;88(9), 4795-802. Anal Chem..PubMedID: 27033327
  • O'Flaherty, Roisin, Harbison, Aoife M., Hanley, Philip J., Fadda, Elisa, Rudd, Pauline, Taron, Chris (2017). Aminoquinoline fluorescent labels obstruct efficient removal of N-glycan corex J Proteome Res. 16, 4237-4243.
Quality Control Assay
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Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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