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α1-2,3,6 Mannosidase

Catalog # Concentration Size List Price Quantity Your Price
P0768S 2000 units/ml 80 units $220.00
$198.00
Catalog # Size List Price Your Price
P0768S 80 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

α1-2,3,6 Mannosidase is a broad specificity exoglycosidase that catalyzes the hydrolysis of terminal, non-reducing α1-2, α1-3 and α1-6 linked mannose 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

α1-2,3,6 Mannosidase, cloned from Jack Bean, and also known as JBM, is a broad specificity exoglycosidase that catalyzes the hydrolysis of terminal α1-2, α1-3 and α1-6 linked mannose residues from oligosaccharides. α1-2,3,6 Mannosidase has a slight preference for α1-2 mannose residues over α1-3 and α1-6 mannose residues.

Substrate Specificity
Product Source
Cloned from Canavalia ensiformis (Jack Bean) and expressed in Pichia pastoris.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  α1-2,3,6 Mannosidase P0768SVIAL 4 1 x 0.04 ml 2,000 units/ml
  GlycoBuffer 4 B1703SVIAL -20 1 x 1 ml 10 X
  Zinc B0768SVIAL 4 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 mannose from 1 nmol of Man(α1,3)-Man(β1,4)-GlcNAc-7-amino-4-methyl-coumarin (AMC), in 1 hour at 37°C in a total reaction volume of 10 µl.

Reaction Conditions

1X GlycoBuffer 4
Supplement with 1X Zinc
Incubate at 37°C

1X GlycoBuffer 4
50 mM sodium acetate
(pH 4.5 @ 25°C)

Storage Buffer

20 mM Tris-HCl
50 mM NaCl
pH 7.5 @ 25°C

Heat Inactivation

95°C for 10 minutes

Molecular Weight

Theoretical: 110 kDa

Unit Assay Conditions

Two fold dilutions of α1-2,3,6 Mannosidase are incubated with 1 nmol AMC-labeled substrate, 1X GlycoBuffer 4 and 1X Zinc 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).

Companion Products
Notes
  • α1-2,3,6 Mannosidase is a glycosylated protein. The theoretical molecular weight is 110 kDa, with two subunits at 66 kDa and 44 kDa (2).
  • The optimal pH for α1-2,3,6 Mannosidase is 4.5. Enzyme activity is enhanced by the presence of 2 mM Zn2+ in the reaction.
  • In order to achieve complete removal of all non-reducing terminalα-linked mannose residues, it may be necessary to use an increasedenzyme concentration as well as an overnight incubation time (18 hours).
  • To achieve complete removal of α1-6 mannose residues from a complexsubstrate, a sequential digest using α1-2,3,6 Mannosidase followed byα1-6 Mannosidase (NEB #P0727) may be necessary. 
  • Reactions may be scaled-up linearly to accommodate larger reaction volumes.
  • The amount of exoglycosidase enzyme required varies when differentsubstrates are used. Start with 1-2 μl for 1 μg of glycoprotein or 100nM of oligosaccharide for one hour in a 10-25 μl reaction. If there isstill undigested material, let the reaction go overnight.
References
  • Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.
  • Kumar, S.N. et al (2014). Glycobiology. 24, 252-261.
Web Tools
Citations
Additional Citations
  • Degani, G., Barbiroli, A., Magnelli, P., Digiovanni, S., Altomare, A., Aldini, G., & Popolo, L. (2019) Insights into the effects of N-glycosylation on the characteristics of the VC1 domain of the human receptor for advanced glycation end products (RAGE) secreted by Pichia pastoris Glycoconjugate Journal 36-1, 27-38.PubMedID: 30612271, DOI: 10.1007/s10719-018-09855-x
Publications
  • Degani, G., Barbiroli, A., Magnelli, P., Digiovanni, S., Altomare, A., Aldini, G., & Popolo, L. (2019). Insights into the effects of N-glycosylation on the characteristics of the VC1 domain of the human receptor for advanced glycation end products (RAGE) secreted by Pichia pastoris Glycoconjugate Journal. 36-1, 27-38.PubMedID: 30612271, DOI: 10.1007/s10719-018-09855-x
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
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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.

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