Product Class: Other

β1-3,4 Galactosidase
NEBU cloned at NEB recombinant 37 65 Heat

Product Introduction

β1-3,4 Galactosidase is an exoglycosidase that catalyzes the hydrolysis of terminal, non-reducing β1-3 and β1-4-linked galactose 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
Catalog # Size Concentration
P0746S 400.0 units 8000 units/ml
P0746L 2000.0 units 8000 units/ml

Product Information

Description

β1-3,4 Galactosidase, cloned from bovine testis and also known as BTG, is a highly specific exoglycosidase that catalyzes the hydrolysis of terminal β1-3 and β1-4 linked galactose residues from oligosaccharides.
P0746 Specificity Figure

Product Source

Cloned from bovine testis and expressed in Pichia pastoris.
This product is related to the following categories:
Exoglycosidases Products
This product can be used in the following applications:
Glycan Sequencing,
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
  • P0746S     4    
      β1-3,4 Galactosidase P0746SVIAL 4 1 x 0.05 ml 8,000 units/ml
      GlycoBuffer 4 B1703SVIAL -20 1 x 1 ml 10 X
  • P0746L     4    
      β1-3,4 Galactosidase P0746LVIAL 4 1 x 0.25 ml 8,000 units/ml
      GlycoBuffer 4 B1703SVIAL -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 terminal, β-D-galactose from 1 nmol Galβ1-4GlcNAcβ1-3Galβ1-4Glc-7-amino-4-methyl-coumarin (AMC), in 1 hour at 37°C in a total reaction volume of 10 µl.

Reaction Conditions

1X GlycoBuffer 4
Incubate at 37°C

1X GlycoBuffer 4
50 mM sodium acetate
(pH 4.5 @ 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

Theoretical: 71 kDa

Unit Assay Conditions

Two fold dilutions of β1-3,4 Galactosidase are incubated with 1 nmol AMC-labeled substrate and 1X GlycoBuffer 4 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).

Product Notes

  1. The optimal pH for β1-3,4 Galactosidase is 4.5.
  2. Although rare in nature, β1-6 linked galactose residues have been shown to be hydrolyzed by β1-3,4 Galactosidase at a much slower rate than β1-3 or β1-4 linked galactose residues (2).
  3. Reactions may be scaled-up linearly to accommodate larger reaction volumes. 
  4. The amount of exoglycosidase enzyme required varies when different substrates are used. Start with 1-2 μl for 1 µg of glycoprotein or 100 nM of oligosaccharide for one hour in a 10-25 μl reaction. If there is still undigested material, let the reaction go overnight.
  5. β1-3,4 Galactosidase is a glycosylated protein.

References

  1. Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.
  2. Distler, J. and Jourdian, G. (1973). J. Biol. Chem. 248, 6772-6780.

Protocols, Manuals & Usage

Protocols

  1. Typical Reaction Conditions for β1-3,4 Galactosidase Reaction Protocol (P0746)

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is the difference between β1-4Galactosidase, β1-4Galactosidase S and β1-3,4Galactosidase? 
  2. Can β1-3,4Galactosidase cleave β1-6 linked galactose residues? 
  3. What is a good positive control for β1-3,4 Galactosidase?
  4. Can I use β1-3,4 Galactosidase in a double digest with other exoglycosidases and/or endoglycosidases?
  5. What is a good method to re-purify a glycan or glycopeptide after exoglycosidase treatment?
  6. How much exoglycosidase should be used?
  7. What are Glycosidases and their uses?
  8. Does this enzyme require denaturing conditions to act on glycoproteins?
  9. Do detergents inhibit glycosidases?
  10. How much exoglycosidase should be used?

Tech Tips

  • Repeated freeze/thaw cycles may reduce activity. Recommended storage temperature is 4°C
  • The optimal pH for β1-3,4 Galactosidase is 4.5. This differs from the majority of exoglycosidases, which have an optimal pH at 5.5.
  • β1-3,4 Galactosidase releases β1-3 and β1-4 galactose residues with equivalent efficiency
  • Using 1-2 µl is a good starting point for a 1 hr incubation of 1µg of glycoprotein or 100 nM of oligosaccharide.

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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