Product Class: Other

α1-3,4 Fucosidase
NEBU cloned at NEB recombinant 37 65 Heat ralbumin

Product Introduction

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

Catalog # Size Concentration
P0769S 200.0 units 4000 units/ml
P0769L 1000.0 units 4000 units/ml

Product Information

Description

 

Product Source

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

Specificity

This product is related to the following categories:
Exoglycosidases Products,
Proteome Analysis Products,
This product can be used in the following applications:
Expression Systems,
Glycan Sequencing,
Proteomics,
Recombinant Glycoprotein Expression,
Glycoprotein Analysis

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • P0769S     4    
      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
  • P0769L     4    
      α1-3,4 Fucosidase P0769LVIAL 4 1 x 0.25 ml 4,000 units/ml
      GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X
      Purified BSA B9001SVIAL -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

  1. Wong-Madden, S. T. and Landry, D. (1995). Glycobiology. 5, 19-28.

Protocols, Manuals & Usage

Protocols

  1. Typical Reaction Conditions for α1-3, 4 Fucosidase (P0769)

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is the difference between α1-2 Fucosidase, α1-3,4,6 Fucosidase and α1-3,4 Fucosidase?
  2. Can I use α1-3,4 Fucosidase in a double digest with other exoglycosidases and/or endoglycosidases?
  3. Can α1-3,4 Fucosidase cleave core α1-3 fucose residues?
  4. Can α1-3,4 Fucosidase and α1-2,3,4,6 Fucosidase cleave branched and complex substrates?
  5. What are Glycosidases and their uses?
  6. Do detergents inhibit glycosidases?
  7. What is a good method to re-purify a glycan or glycopeptide after exoglycosidase treatment?
  8. Does this enzyme require denaturing conditions to act on glycoproteins?
  9. How much exoglycosidase should be used?

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, Safety & Legal

Quality Assurance Statement

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 DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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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