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β1-3 Galactosidase

Catalog # Concentration Size List Price Quantity Your Price
P0726S 10000 units/ml 500 units $220.00
$198.00
Catalog # Size List Price Your Price
P0726S 500 units $220.00
$198.00
Catalog #
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

β1-3 Galactosidase is a highly specific exoglycosidase that catalyzes the hydrolysis of β1-3 and, at a much lower rate, β1-6 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 24 months
Substrate Specificity:

β1-3 Galactosidase is a highly specific exoglycosidase that catalyzes the hydrolysis of β1-3 and, at a much lower rate, β1-6 linked D-galactopyranosyl residues from oligosaccharides. The approximate kinetic data show > 100-fold preference for β1-3 over β1-6 linkages (1,2), and > 500-fold preference from β1-3 over β1-4 linkages (3). 

Detailed Specificity: The GlcNAc(β1-6) residue is the only anomeric configuration that can effect the specificity of the enzyme enabling cleavage of the non-reducing β1-4Galactose (Fig. 1).


 Figure 1: 
Figure 1


Selling concentration of the enzyme will cut the β1-4Galactose linkage as shown in (A) due to the adjacent GlcNAcβ1-6 anomer. This cleavage will not occur if the selling concentration of the enzyme is diluted 16-fold, as shown in (B).

Product Source
Cloned from Xanthomonas manihotis and expressed in E. coli.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  β1-3 Galactosidase P0726SVIAL -20 1 x 0.05 ml 10,000 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 β-D-galactose from 1 nmol of Galβ1-3GlcNAcβ1-3Galβ1-4Glc-7-amino-4-methyl-coumarin (AMC), in 1 hour at 37°C in a total reaction volume of 10 µl.

Reaction Conditions

1X GlycoBuffer 1
Supplement with 100 µg/µl 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

50 mM NaCl
20 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C

Heat Inactivation

65°C for 10 minutes

Molecular Weight

Theoretical: 66 kDa

Unit Assay Conditions

Two fold dilutions of β1-3 Galactosidase are incubated with 1 nmol AMC-labeled substrate in 1X GlycoBuffer 1, supplemented with 100 µg/ml rHSA, in a 10 µl reaction. The reaction mix is incubated for 1 hour at 37°C. Separation of reaction products are visualized via thin layer chromatography (1).

Companion Products
Notes
  • Optimal incubation times and enzyme concentrations must be determined empirically for a particular substrate.
References
  • Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.
  • Guthrie, E.P. and Taron, C.H. New England Biolabs. Unpublished observation
  • Monks, B. New England Biolabs. Unpublished observation
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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