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α1-3,4 Fucosidase

Catalog # Concentration Size List Price Quantity Your Price
P0769S 4000 units/ml 200 units $217.00
$195.30
Catalog # Size List Price Your Price
P0769S 200 units $217.00
$195.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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

• 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

 
Product Source
Cloned from the sweet almond tree (Prunus dulcis) and expressed in Pichia pastoris.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  α1-3,4 Fucosidase P0769SVIAL 4 1 x 0.05 ml 4,000 units/ml
  GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X
  Recombinant Albumin (Low-glycerol) B9203SVIAL -20 1 x 0.25 ml 10 mg/ml

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave > 95% of the α-fucose from 1 nmol of Galβ1-4GlcNAcβ1-3(Fucα1-3)Galβ1- 4Glc-7-amino-4-methyl-coumarin (AMC), in 1 hour at 37°C in a total reaction volume of 10 μl

Reaction Conditions

1X GlycoBuffer 1
Supplement with 100 µg/ml Recombinant Albumin (Low-glycerol)
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

65°C for 10 minutes

Molecular Weight

Apparent: 56000 daltons

Unit Assay Conditions

Two fold dilutions of a1-3,4 Fucosidase are incubated with 1 nmol AMC-labeled substrate in 1X GlycoBuffer 1 supplemented with 1X rHSA 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 (1).

References
  • 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. 
    • Higher enzyme concentrations and longer incubation times may be required for the removal of α1-3 and α1-4 fucose residues from complex N-glycans. 
    • α1-3,4 Fucosidase can cleave the α1-3 fucose on the outer arm of N-linked glycans but is unable to remove the core α1-3 fucose residue. 
    • The storage buffer does not contain rHSA; however, we recommend supplementing the reaction with rHSA for optimal enzyme activity.
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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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