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β-N-Acetylhexosaminidasef

Catalog # Concentration Size List Price Quantity Your Price
P0721S 5000 units/ml 500 units $115.00
$103.50
Catalog # Size List Price Your Price
P0721S 500 units $115.00
$103.50
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

β-N-Acetylhexosaminidasef is an exoglycosidase that catalyzes the hydrolysis of terminal, non-reducing β-N-Acetylgalactosamine and glucosamine residues from oligosaccharides.

  • Can be used for Epigenetic applications to remove O-GlcNAc from serine/threonine residues on transcription factors, histones, RNA-binding proteins and other O-GlcNAc modified 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
β-N-Acetylhexosaminidasef is a recombinant protein fusion of β-N-Acetylhexosaminidase (1) and maltose binding protein. It has identical activity to β-N-Acetylhexosaminidase. β-N-Acetylhexosaminidasef catalyzes the hydrolysis of terminal β-D-N-acetyl-galactosamine and glucosamine residues from oligosaccharides.  β-N-Acetylhexosaminidasef can be used to remove O-GlcNAc (N-Acetylglucosaminidase on serine/threonine)(4). 

Substrate Specificity:
Highlights
  • Epigenetics applications:  O-GlcNac (N-Acetylhexosamine attached to serine or threonine) is a regulator of nuclear protein function. β-N-Acetylhexosaminidasef removes O-GlcNAc from transcription factors, histones, RNA-binding proteins, etc.
Product Source
Cloned from Streptomyces plicatus and overexpressed in E. coli.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  β-N-Acetylhexosaminidasef P0721SVIAL -20 1 x 0.1 ml 5,000 units/ml
  GlycoBuffer 1 B1727SVIAL -20 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 β-D-N-Acetylgalactosamine from 1 nmol of GalNAcβ1-4Galβ1-4Glc-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
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
5 mM EDTA
pH 7.5 @ 25°C

Heat Inactivation

75°C for 10 minutes

Molecular Weight

Theoretical: 100000 daltons

Terminal Monosacaride

  • N-Acetyl-galactosamine and N-Acetyl-glucosamine Residues

Unit Assay Conditions

Two fold dilutions of β-N-Acetyl-hexosaminidasef are incubated with 1 nmol AMC-labeled substrate in 1X GlycoBuffer 1 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 (3).

References
  • Robbins, P. et al. (1992). Gene. 111, 69-76.
  • Guan, C. and Wong, S. New England Biolabs.. Unpublished observation
  • Wong-Madden, S.T. and Landry D. (1995). Glycobiology. 5, 19-28.
  • Magnelli, P.E. et al Methods Molecular Biology. 801, 189-211.
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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