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α1-3,6 Galactosidase

Catalog # Concentration Size List Price Quantity Your Price
P0731S 4000 units/ml 100 units $219.00
$197.10
Catalog # Size List Price Your Price
P0731S 100 units $219.00
$197.10
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

α1-3,6 Galactosidase is a highly specific exoglycosidase that catalyzes the hydrolysis of α1-3 and α1-6 linked galactose residues from oligosaccharides.
• Removal of the antigenic Galα1,3Gal epitope from glycoproteins
• 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

 α1-3,6 Galactosidase is a highly specific exoglycosidase that catalyzes the hydrolysis of α1-3 and α1-6 linked D-galactopyranosyl residues from oligosaccharides. 



Detailed Specificity: Specificity can vary depending on incubation time and branching structure.

Figure 1: Detailed specificity of α1-3,6 Galactosidase

Figure 1: Detailed specificity of α1-3, 6 Galactosidase
Reaction (B) contained 4 units of α1-3, 6 Galactosidase, 1X GlycoBuffer 1 and 1X rHSA in a total reaction volume of 10 µl. Reaction (B) shows that branched fucose inhibits cleavage. Reaction (A) contained 24 units of α1-3, 6 Galactosidase and 100 units of Neuraminidase, followed by a heat kill at 65°C for 10 minutes and a 2 hour digestion with 16 units of β1-4 Galactosidase. The reaction in (B) contained 1X GlycoBuffer 1 and 1X BSA in a total reaction volume of 20 µl. The reactions were incubated at 37°C. Complete digestion of the α1-3, 6 Galactosidase was determined by an observation of complete transformation of the substrate in (A) to the non-reducing terminal N-acetylglucosamine tetra antennary oligosaccharide.
Highlights
Removal of the antigenic Galα1,3Gal epitope from glycoproteins.
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
  α1-3,6 Galactosidase P0731SVIAL 4 1 x 0.025 ml 4,000 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 for 10 minutes

Molecular Weight

Apparent: 70000 daltons

Unit Assay Conditions

Two fold serial dilutions of α1-3,6 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 for 1 hour at 37°C. Separation of reaction products are visualized via thin layer chromatography (1).

Notes
  • Avoid repeated freeze-thaw cycles.
References
  • Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.
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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