Product Class: Other

α1-6 Mannosidase
NEBU cloned at NEB recombinant 37 65 Heat ralbumin

Product Introduction

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
Catalog # Size Concentration
P0727S 800.0 units 40000 units/ml
P0727L 4000.0 units 40000 units/ml

Product Information

Description

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.
This product is related to the following categories:
Exoglycosidases Products
This product can be used in the following applications:
Recombinant Glycoprotein Expression

Reagents Supplied

Reagents Supplied

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).

Product Notes

  1. p-nitrophenyl-α-D­­­-mannopyranoside is NOT a substrate for this enzyme.
  2. Avoid repeated freeze-thaw cycles.

References

  1. Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.

Protocols, Manuals & Usage

Protocols

  1. Typical Reaction Conditions for α1-6 Mannosidase (P0727)

Usage & Guidelines

FAQs & Troubleshooting

FAQs

  1. What is a good positive control for α1-6 Mannosidase?
  2. How much exoglycosidase should be used?
  3. Why have the NEB Glycosidase enzymes changed reaction buffers? What are the new reaction buffers and can I still use an enzyme with its old buffer? Where can I find the composition of the old buffers?
  4. Do detergents inhibit exoglycosidases/endoglycosidases?
  5. What are Glycosidases and their uses?
  6. How much exoglycosidase should be used?

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, Safety & Legal

Quality Assurance Statement

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 DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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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