Product Class: Other

α1-3,4,6 Galactosidase
NEBU cloned at NEB recombinant 37 65 Heat ralbumin

Product Introduction

α1-3,4,6 Galactosidase is a broad specificity exoglycosidase that catalyzes the hydrolysis of terminal, non-reducing α1-3, α1-4 and α1-6-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
P0747S 200.0 units 8000 units/ml
P0747L 1000.0 units 8000 units/ml

Product Information

Description

 

Product Source

Cloned from green coffee bean and expressed in E. coli.

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
  • P0747S     4    
      α1-3,4,6 Galactosidase P0747SVIAL 4 1 x 0.025 ml 8,000 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
  • P0747L     4    
      α1-3,4,6 Galactosidase P0747LVIAL 4 1 x 0.125 ml 8,000 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 terminal, α-D-galactose from 1 nmol Galα1-3Galβ1-4Gal- 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

Molecular Weight

Theoretical: 39.7 kDa

Specific Activity

71,000 units/mg

Unit Assay Conditions

Two fold dilutions of α1-3,4,6 Galactosidase are incubated with 1 nmol AMC labeled substrate in 1X GlycoBuffer 1 and 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).

Product Notes

  1. 8 units of enzyme wereinactivated by incubation at 65°C for 10 minutes.

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, 6 Galactosidase (P0747)

Application Notes

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. What are Glycosidases and their uses?
  2. What is the difference between α1-3,6 Galactosidase and α1-3,4,6 Galactosidase?
  3. Can I use α1-3,4,6 Galactosidase in a double digest with other exoglycosidases and/or endoglycosidases?
  4. What is a good positive control for α1-3,4,6 Galactosidase?
  5. Does this enzyme require denaturing conditions to act on glycoproteins?
  6. What is a good method to re-purify a glycan or glycopeptide after exoglycosidase treatment?
  7. Do detergents inhibit glycosidases?
  8. How much exoglycosidase should be used?
  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.
  • A good positive control for α1-3,4 6 Galactosidase is p-Nitrophenyl α-D-galactopyranoside
  • 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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