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
Product Information
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.
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
P0721S
-20
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: 100 kDa
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 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 & Change Notifications
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]
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]
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.
