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α1-2,4,6 Fucosidase O

Catalog # Concentration Size List Price Quantity Your Price
P0749L 2000 units/ml 400 units $870.00
$783.00
P0749S 2000 units/ml 80 units $219.00
$197.10
Catalog # Size List Price Your Price
P0749L 400 units $870.00
$783.00
P0749S 80 units $219.00
$197.10
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

α1-2,4,6 Fucosidase O is a broad specificity exoglycosidase that catalyzes the hydrolysis of terminal α1-2, α1-4 and α1-6 linked fucose residues from oligosaccharides. α1-2,4,6 Fucosidase O cleaves α1-6 fucose residues more efficiently than other linkages.

  • 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
α1-2,4,6 Fucosidase O is a broad specificity exoglycosidase that catalyzes the hydrolysis of terminal α1-2, α1-4 and α1-6 linked fucose residues from oligosaccharides. α1-2,4,6 Fucosidase O cleaves α1-6 fucose residues more efficiently than other linkages.
• Recombinant enzyme with no detectable endoglycosidase or other exoglycosidase contaminating activities
• Glycerol-free foroptimal 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 Omnitrophica bacterium and expressed in E. coli.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  α1-2,4,6 Fucosidase O P0749SVIAL 4 1 x 0.04 ml 2,000 units/ml
  GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X
  α1-2,4,6 Fucosidase O P0749LVIAL 4 1 x 0.2 ml 2,000 units/ml
  GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave > 95% of the fucose from 1 nmol of G0F from human IgG [GlcNAcβ1-2Manα1-6(GlcNAcβ1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc(Fucα1-6)-AMAC], in 1 hour at 37°C in a total reaction volume of 10 µl.

Reaction Conditions

1X GlycoBuffer 1
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 @ 20°C

Heat Inactivation

65°C for 10 minutes

Molecular Weight

Apparent: 49 kDa

Unit Assay Conditions

Two fold dilutions of α1-2,4,6 Fucosidase O are incubated with 1 nmol AMAC-labeled substrate and 1X GlycoBuffer 1 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).

Companion Products
Notes
  • The enzymatic unit definition is calculated using a 37°C reaction temperature, to accommodate for simultaneous glycosidase digestions and N-glycan sequencing protocols. However, the optimal reaction temperature for α1-2,4,6 Fucosidase O is 50°C. A 37°C reaction temperature results in ~35% lower activity.
  • In order to achieve complete removal of core α1-6 fucose residues from glycans with instant labels, it may be necessary to use longer incubation times (18 hours).
  • α1-2,4,6 Fucosidase O is only active on an intact antibody if it is trimmed to the trimannosyl core.
  • Reactions may be scaled-up linearly to accommodate larger reaction volumes.
References
  • Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.
Web Tools
Citations
Additional Citations
  • Vainauskas, S., Kirk, C.H., Petralia, L., Guthrie, E.P., McLeod, E., Bielik, A., Luebbers, A., Foster, J.M., Hokke, C.H., Rudd, P.M., Shi, X., Taron, C.H (2018) A novel broad specificity fucosidase capable of core a1-6 fucose release from N-glycans labeled with urea-linked fluorescent dyes Sci RepPubMedID: 29934601, DOI: 1038/s41598-018-27797-0
Publications
  • 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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