O-Glycosidase

Catalog #	Concentration	Size	List Price	Quantity	Your Price
P0733L	40000000 units/ml	10000000 units	$919.00		$827.10
P0733S	40000000 units/ml	2000000 units	$232.00		$208.80

Catalog #	Size	List Price	Your Price
P0733L	10000000 units	$919.00	$827.10
P0733S	2000000 units	$232.00	$208.80

O-Glycosidase, also known as Endo-α-N-Acetylgalactosaminidase, catalyzes the removal of Core 1 and Core 3 O-linked disaccharides from glycoproteins.

Product Source

Cloned from Enterococcus faecalis and expressed in E. coli (1).

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

P0733S -20

O-Glycosidase	P0733SVIAL	-20	1 x 0.05 ml	4.0E7 units/ml
GlycoBuffer 2	B3704SVIAL	-20	1 x 1 ml	10 X
Glycoprotein Denaturing Buffer	B1704SVIAL	-20	1 x 1 ml	10 X
NP-40	B2704SVIAL	-20	1 x 1 ml	10 %

P0733L -20

O-Glycosidase	P0733LVIAL	-20	1 x 0.25 ml	4.0E7 units/ml
GlycoBuffer 2	B3704SVIAL	-20	1 x 1 ml	10 X
Glycoprotein Denaturing Buffer	B1704SVIAL	-20	1 x 1 ml	10 X
NP-40	B2704SVIAL	-20	1 x 1 ml	10 %

Application Features

Removal of Core 1 and Core 3 O-linked disaccharide glycans from glycoproteins

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to remove 0.68 nmol of O-linked disaccharide from 5 mg of neuraminidase digested, non-denatured fetuin (2) in 1 hour at 37°C in a total reaction volume of 100 µl (1 unit of both O-Glycosidase and PNGase F will remove equivalent molar amounts of O-linked disaccharides and N-linked oligosaccharides, respectively).

1X Glycoprotein Denaturing Buffer
0.5% SDS
40 mM DTT

1X NP-40
1% NP-40 in MilliQ-H₂O

Reaction Conditions

1X GlycoBuffer 2
Incubate at 37°C

1X GlycoBuffer 2
50 mM Sodium Phosphate
(pH 7.5 @ 25°C)

Storage Buffer

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

Heat Inactivation

65°C for 10 minutes

Molecular Weight

Apparent: 147000 daltons

Unit Assay Conditions

Two fold serial dilutions of O-Glycosidase are added to a reaction mixture of 5 mg of neuraminidase digested fetuin with 1X GlycoBuffer 2. The reaction mix is then incubated at 37°C for 1 hour. O-linked disaccharide carbohydrates are determined by the Morgan and Elson Assay (1).

Companion Products

Notes

Since O-Glycosidase is inhibited by SDS, it is essential to have NP-40 in the reaction mixture. It is not known why this non-ionic detergent counteracts the SDS inhibition at the present time. Double digest with Endo H must have NP-40 present (NP-40 does not inhibit Endo H).
To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required.
It is necessary to treat glycoproteins concomitantly with Neuraminidase and O-Glycosidase. Neuraminic Acid residues must be removed in order to allow O-Glycosidase to cleave the O-linked disaccharides. A general Neuraminidase (#P0720) or the O-Glycosidase and Neuraminidase bundle (#E0540S) is recommended.
Under denaturing conditions the enzyme activity is increased two-fold. This observation is substrate dependent.

References

Morgan, W.T.J. and Elson, L.A. (1994). Biochem. J.. 28, 988-995.

Protocols

Selection Charts

Endoglycosidase Selection Chart

FAQs

Tech Tips

Don’t forget to include Neuraminidase (P0720) in your reaction! It is necessary to treat glycoproteins concomitantly with Neuraminidase and O-Glycosidase. Neuraminic Acid residues must be removed in order to allow O-Glycosidase to cleave the O-linked disaccharides.

NEB’s O-Glycosidase is the only available O-Glycosidase that catalyzes the removal of Core 1 and Core 3 O-linked disaccharides from glycoproteins

You can use this enzyme under native or denaturing conditions

Under native conditions we recommend adding more enzyme and using longer incubation times

Under denaturing conditions the enzyme activity is increased two-fold. However, this observation may be substrate dependent.

Citations

See more details on Bioz

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]

P0733S_L_v1

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

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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