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β1-3,4 Galactosidase

Catalog # Concentration Size List Price Quantity Your Price
P0746S 8000 units/ml 400 units $220.00
$198.00
Catalog # Size List Price Your Price
P0746S 400 units $220.00
$198.00
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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
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.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  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

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).

Companion Products
Notes
  • The optimal pH for β1-3,4 Galactosidase is 4.5.
  • 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).
  • Reactions may be scaled-up linearly to accommodate larger reaction volumes. 
  • 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.
  • β1-3,4 Galactosidase is a glycosylated protein.
References
  • Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.
  • Distler, J. and Jourdian, G. (1973). J. Biol. Chem. 248, 6772-6780.
Web Tools
FAQs
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 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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