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
Product Information
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
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.
This product is related to the following categories:
The following reagents are supplied with this product:
NEB #
Component Name
Component #
Stored at (°C)
Amount
Concentration
P0731S
4
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).
Product Notes
Avoid repeated freeze-thaw cycles.
References
Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.
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