Product Class: Other

Endo-β-Galactosidase
Gray property icon for NEBU Gray property icon for cloned at NEB recombinant Gray property icon for incubation temperature 37° 55 Heat

Product Introduction

  • Removal of poly-N-acetyllactosamine in glycoproteins and glycolipids
  • Broader substrate specificity and higher activity compared to other enzymes from Escherichia freundii and Bacteroides fragilis
  • 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
 
Bioz Badge Exists : True
Catalog # Size Concentration
P0777S 50 µg 0.5 mg/ml

Product Information

Description

Endo-β-Galactosidase is an endoglycosidase that hydrolyzes internal β(1-4)-galactose linkages within a variety of sulfated and nonsulfated poly-N-acetyllactosamine structures, [Galβ(1-4)-↓-GlcNAcβ(1-3)], derived from milk, glycoproteins and glycolipids. This enzyme is used for structural and functional analysis of poly-N-acetyllactosamine in glycoconjugates.

This endo-beta-galactosidase (endo-b-galactosidase) enzyme from F. keratolyticus exhibits broader substrate specificity and higher catalytic efficiency compared to others from E. freundii and B. fragilis.

EBG is highly specific to repeating Gal-GlcNAc (polyLacNAc) units. PolyLacNAc occurs in N-glycans, O-glycans, and glycolipids, thus it is not only a cancer biomarker (e.g., breast, colon), but also widely present as a glycan epitope involved in immunology, developmental biology, and disease.

Specificity:

Mechanism of Endo-B-galactosidase

null

Product Source

Cloned from Flavobacterium keratolyticus and expressed in E. coli
This product is related to the following categories:
Endoglycosidases
This product can be used in the following applications:
Glycoprotein Analysis

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • NEB # Stored at (°C)
    P0777S -20
    Component Name Component # Stored at (°C) Amount Concentration
    Endo-β-Galactosidase P0777SVIAL -20 1 x 50 µg 0.5 mg/ml
    GlycoBuffer 3 B1720SVIAL -20 1 x 1 ml 10 X

Properties & Usage

Unit Definition

The enzyme activity is determined by its ability to cleave over 95% of 2 nmol Galβ1-4GlcNAcβ1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glc-7-amino-4-methyl-coumarin (AMC) using less than 10 ng of enzyme within 1 hour at 37°C, in a total reaction volume of 20 μl.

Reaction Conditions

1X GlycoBuffer 3
Incubate at 37°C

1X GlycoBuffer 3
50 mM sodium acetate
(pH 6 @ 25°C)

Storage Buffer

50 mM sodium acetate
50 mM NaCl
pH 5.5 @ 25°C

Heat Inactivation

55°C for 10 minutes

Molecular Weight

Apparent: 40.8 kDa

Unit Assay Conditions

Two fold dilutions of Endo-β-Galactosidase are incubated with 2 nmol AMC labeled substrate in 1X GlycoBuffer 3 in a 20 μl reaction. The reaction mix is incubated at 37°C for 1 hour. Separation of reaction products are visualized via thin layer chromatography.

Features

  • This endo-beta-galactosidase (endo-b-galactosidase) enzyme from F. keratolyticus exhibits broader substrate specificity and higher catalytic efficiency compared to others from E. freundii and B. fragilis.
  • The enzyme is active in a wide range of pH conditions (pH 3.5 to pH 9.0).
  • The enzyme works at 37°C and below.

This enzyme is ideal for:

  • Removal of poly-N-acetyllactosamine in glycoproteins and glycolipids.
  • Structural and functional analysis of poly-N-acetyllactosamine in glycoconjugates.
  • Researchers studying polyLacNAc in mammalian systems
  • Those using B. fragilis EBG, which tend to have lower activity
  • Cell surface studies (analogous to PNGase F)
  • Analysis of polyLacNAc-binding proteins (e.g., Galectins)
  • Glycopeptide MS workflows (to improve ionization and fragmentation; in combination with PNGaseF)
  • Comparative studies (+/− polyLacNAc)
  • Immunology, pathology, and stem cell differentiation 

FAQs & Troubleshooting

FAQs

  1. Do detergents inhibit Endo- β-Galactosidase (NEB #P0777)?
  2. How much Endo-β-Galactosidase (NEB #P0777) should be used in a reaction?
  3. Can I use a different buffer than GlycoBuffer 3 supplied with Endo-β-Galactosidase (NEB #P0777)?
  4. What if I have incomplete digestion when using Endo-β-Galactosidase (NEB #P0777)?
  5. What is the difference between Endo-β-Galactosidase, β1-4 Galactosidase, β1-4 Galactosidase S and β1-3,4 Galactosidase?

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.