α1-6 Mannosidase is a highly specific exoglycosidase that removes unbranched α1-6 linked mannose residues from oligosaccharides.
When used in conjunction with α1-2,3 Mannosidase, the α1-6 Mannosidase will cleave α1-6 mannose residues from branched carbohydrate substrates.
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
Substrate Specificity:
α1-6 Mannosidase is a highly specific exoglycosidase that removes unbranched α1-6 linked D-mannopyranosyl residues from oligosaccharides (1,2). When used in conjunction with α1-2,3 Mannosidase, the α1-6 Mannosidase will cleave α1-6 Mannose residues from branched carbohydrate substrates.
Figure 1: Detailed specificity of α1-6 Mannosidase All reactions contained 32 units of α1-2,3 Mannosidase (NEB #P0729 ), 40 units of α1-6 Mannosidase, 1X GlycoBuffer 1 and 1X rHSA In a total reaction volume of 10 µl. Reactions were incubated at 37°C. The substrate depicted in (E) will not cut to completion. If this structure exists in any substrate it will be impervious to cleavage by α1-6 Mannosidase. Note: When used alone, α1-6 Mannosidase will still act only on linear substrates. When used in conjunction with α1-2,3 Mannosidase, the α1-6 Mannosidase will cleave α1-6 Mannose residues from branched carbohydrate substrates.
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
P0727S
4
α1-6 Mannosidase
P0727SVIAL
4
1 x 0.02 ml
40,000 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
P0727L
4
α1-6 Mannosidase
P0727LVIAL
4
1 x 0.1 ml
40,000 units/ml
GlycoBuffer 1
B1727SVIAL
-20
1 x 1 ml
10 X
Purified BSA
B9001SVIAL
-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-mannose from 1 nmol of Manα1-6Manα1-6Man-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
50 mM NaCl
20 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C
Heat Inactivation
65°C for 10 minutes
Molecular Weight
Apparent: 51 kDa
Unit Assay Conditions
Two fold dilutions of α1‑6 Mannosidase 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).
Using 1-2 µl is a good starting point for a 1 hr incubation of 1µg of glycoprotein or 100 nM of oligosaccharide.
When used alone α1-6Man removes only unbranched α1-6 linked mannose residues from oligosaccharides.
p-nitrophenyl-a-D-Mannopyranoside (pNP-Man) is NOT a substrate for this enzyme.
Repeated freeze/thaw cycles may reduce activity. Recommended storage temp 4°C.
Citations & Technical Literature
Citations
Product Citation Tool
Quality, Safety & Legal
Quality Control Assays
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Specifications & Change Notifications
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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]
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