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α1-6 Mannosidase

Catalog # Concentration Size List Price Quantity Your Price
P0727S 40000 units/ml 800 units $220.00
$198.00
Catalog # Size List Price Your Price
P0727S 800 units $220.00
$198.00
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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
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
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.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  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

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 α1­­­­­­­­â€‘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).

Companion Products
Notes
  • p-nitrophenyl-α-D­­­-mannopyranoside is NOT a substrate for this enzyme.
  • Avoid repeated freeze-thaw cycles.
References
  • Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.
FAQs
Tech Tips

  • 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.

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
Legal And Disclaimer

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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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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