α-N-Acetylgalactosaminidase is an exoglycosidase that catalyzes the hydrolysis of terminal, non-reducing α- N-acetylgalactosamine residues from oligosaccharides and N-glycans attached to proteins.
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
Product Information
α-N-Acetylgalactosaminidase is a highly specific exoglycosidase that catalyzes the hydrolysis of α-linked D-N-acetylgalactosamine residues from oligosaccharides and N-glycans attached to proteins.
Substrate Specificity:
Product Source
Cloned from Chryseobacterium meningosepticum 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
P0734S
-20
α-N-Acetylgalactosaminidase
P0734SVIAL
-20
1 x 0.15 ml
20,000 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-N-acetylgalactosamine from 1 nmol (GalNAcα1-3)(Fucα1-2)Galβ1-4Glc-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/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: 47 kDa
Unit Assay Conditions
Two fold dilutions of α-N-Acetylgalactosamininidase 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).
Product 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.
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Specifications & Change Notifications
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