β-N-Acetylglucosaminidase S is an exoglycosidase that catalyzes the hydrolysis of terminal, non-reducing β-N-Acetylglucosamine 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
Product Information
β-N-Acetylglucosaminidase S is
a highly specific exoglycosidase that catalyzes
the hydrolysis of terminal, non-reducing β-N-Acetylglucosamine
residues from oligosaccharides.
Specificity Detailed Specificity
β-N-Acetylglucosaminidase S is able to efficiently cleave bisecting β-N-Acetylglucosaminidase residues.
Product Source
Cloned from Streptococcus pneumoniae 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
P0744S
4
β-N-Acetylglucosaminidase S
P0744SVIAL
4
1 x 0.025 ml
4,000 units/ml
GlycoBuffer 1
B1727SVIAL
-20
1 x 1 ml
10 X
P0744L
4
β-N-Acetylglucosaminidase S
P0744LVIAL
4
1 x 0.125 ml
4,000 units/ml
GlycoBuffer 1
B1727SVIAL
-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, non-reducing β-N-Acetylglucosamine from 1 nmol GlcNAcβ1-4GlcNAcβ1-4GlcNAc-7-amino-4-methylcoumarin (AMC), in 1 hour at 37°C in a total reaction volume of 10 μl.
Reaction Conditions
1X GlycoBuffer 1
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
No
Molecular Weight
Apparent: 125 kDa
Unit Assay Conditions
Two fold dilutions of β-N-Acetylglucosaminidase S are incubated with 1 nmol AMC-labeled substrate and 1X GlycoBuffer 1 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).
Product Notes
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.
β-N-Acetylglucosaminidase S cannot be heat inactivated.
References
Wong-Madden, S. T. and Landry, D. (1995). Glycobiology. 5, 19-28.
Repeated freeze/thaw cycles may reduce activity. Recommended storage temperature is 4°C.
Using 1-2 µl is a good starting point for a 1 hr incubation of 1µg of glycoprotein or 100 nM of oligosaccharide.
A good positive control for β-N-Acetylglucosaminidase S is pNP-GlcNAc.
β-N-Acetylglucosaminidase S is not a general ß-hexosamindase, it is instead a highly specific exoglycosidase that catalyzes the hydrolysis of all linkages of terminal, non-reducing ß-N-acetylglucosamine (ß-GlcNAc) residues from oligosaccharides.
β-N-Acetylglucosaminidase S is able to efficiently cleave bisecting β-N-Acetylglucosamine residues.
β-N-Acetylglucosaminidase S cannot be heat inactivated.
Citations & Technical Literature
Citations
Product Citation Tool
Publications
Albrecht, S., Vainauskas, S., Stöckmann, H., McManus, C., Taron, C.H. and Rudd, P.M. (2016) Comprehensive Profiling of Glycosphingolipid Glycans Using a Novel Broad Specificity Endoglycoceramidase in a High-Throughput Workflow. Anal Chem; May 3;88(9), 4795-802. Anal Chem.. PubMedID: 27033327
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Specifications & Change Notifications
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