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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  • Vainauskas, S., New England Biolabs, Inc. Unpublished observation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
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
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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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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