O-Glycosidase

Catalog #	Concentration	Size	List Price	Your Price
P0733S	40,000,000 units/ml	2,000,000 units	$135.00	Sign In Sign In
P0733L	40,000,000 units/ml	10,000,000 units	$539.00	Sign In Sign In

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

Substrate Specificity:

Product Source
Cloned from Enterococcus faecalis and expressed in E. coli (1).
Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration

Glycoprotein Denaturing Buffer -20 10 X

NP-40 -20 10 %

GlycoBuffer 2 -20 10 X

Product Categories:

Endoglycosidases Products,

Proteome Analysis Products

Applications:

Expression Systems,

Protein Digestion,

Glycan Sequencing
,
Glycoprotein Analysis
,
Recombinant Glycoprotein Expression
- Advantages and Features
  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).
  
  Non-Denaturing Unit Definition Assay
  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 (2).
  
  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 Temperature
  -20°C
  Storage Conditions
  
  20 mM Tris-HCl
  50 mM NaCl
  1 mM EDTA
  pH 7.5 @ 25°C
  
  Heat Inactivation
  65°C for 10 min
  Molecular Weight
  Apparent: 147000 daltons
- Related Products
  Companion Products
  
  O-Glycosidase & Neuraminidase Bundle
  
  Endo H
  
  Endo H_f
  
  PNGase F
  
  PNGase F (Glycerol-free)
  
  α2-3,6,8 Neuraminidase
- Product 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
  Koutsioulis, D., Landry, D. and Guthrie, E.P. (2008). Glycobiology. 18, 799-805.
  
  Morgan, W.T.J. and Elson, L.A. (1994). Biochem. J.. 28, 988-995.
Protocols, Manuals & Usage
- Protocols
  O-Glycosidase Application Note 1 (P0733)
  
  O-Glycosidase (P0733)
  
  Endo-α-N-Acetylgalactosaminidase Application Note 1
- Application Notes
  Enzymatic deglycosylation of a protein containing Core 1 O-glycans with O-Glycosidase
  
  Analysis of a Fusion Protein using the Protein Deglycosylation Mix II and Mass Spectrometry
- Datacards
  The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)
  
  P0733Datasheet-Lot0011206
  
  P0733Datasheet-Lot0011210
  
  P0733Datasheet-Lot0011302
  
  P0733Datasheet-Lot0011310
  
  P0733Datasheet-Lot0031403
  
  P0733Datasheet-Lot0031408
  
  P0733Datasheet-Lot0031412
  
  P0733Datasheet-Lot0011504
  
  P0733Datasheet-Lot0011505
  
  P0733Datasheet-Lot0031507
Tools & Resources
- Selection Charts
  Endoglycosidase Selection Chart
FAQs & Troubleshooting
- FAQs
  What are the typical reaction conditions for O-Glycosidase?
  
  I tried using O-Glycosidase on my glycoprotein and didn't see removal of the carbohydrate. What could be the problem?
  
  How much O-Glycosidase should I use to remove my carbohydrate under native conditions?
  
  Do detergents inhibit O-Glycosidase?
  
  Can Neuraminidase be used together in a digest with PNGase F and O-Glycosidase?
  
  Can I double digest PNGase F and O-Glycosidase?
  
  What is the difference between PNGase F, Endo H and O-Glycosidase?
  
  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?
  
  What are glycosidases and their uses?
  
  Is it necessary to treat my glycoprotein concomitantly with Neuraminidase and O-Glycosidase?
  
  How do I inhibit O-Glycosidase?
- 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 & Technical Literature
- Brochures
  Glycoproteomics Brochure
  
  Glycoproteomics Technical Guide
- Citations
  
  Product Citation Tool
  
  Powered by Bioz See more details on Bioz
- Feature Articles
  Using Glycosidases to Remove Trim or Modify Glycans on Therapeutic Protein
Quality, Safety & Legal
- Quality Control Assays
  
  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]
  
  P0733S_L_v1_0031603
  
  P0733S_L_v1_0031612
  
  P0733S_L_v1_0031706
  
  P0733S_L_v1_0031801
  
  P0733S_v1_10011712
  
  P0733S_v1_10015197
  
  P0733L_v1_10025560
  
  P0733S_v1_10027227
  
  P0733S_v1_10033432
  
  P0733S_v1_10039238
  
  P0733L_v1_10043512
  
  P0733S_v1_10043510
  
  P0733L_v1_10050882
  
  P0733S_v1_10051572
  
  P0733S_v1_10054778
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  O-Glycosidase
  
  Glycoprotein Denaturing Buffer
  
  NP-40
  
  GlycoBuffer 2
- Datacards
  The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)
  
  P0733Datasheet-Lot0011206
  
  P0733Datasheet-Lot0011210
  
  P0733Datasheet-Lot0011302
  
  P0733Datasheet-Lot0011310
  
  P0733Datasheet-Lot0031403
  
  P0733Datasheet-Lot0031408
  
  P0733Datasheet-Lot0031412
  
  P0733Datasheet-Lot0011504
  
  P0733Datasheet-Lot0011505
  
  P0733Datasheet-Lot0031507
- Legal and Disclaimers
  
  This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
  
  While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
  
  For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
  
  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.

Applications

Product Categories

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Tools

Resources

Applications

Product Categories

You’ll be Thrilled to Pieces

Tools

Resources

O-Glycosidase

Substrate Specificity:

Product Source

Reagents Supplied

Application Features

Unit Definition

Reaction Conditions

Storage Temperature

Storage Conditions

Heat Inactivation

Molecular Weight

Companion Products

Product Citation Tool

Other Products You May Be Interested In

O-Glycosidase & Neuraminidase Bundle

Protein Deglycosylation Mix II

α2-3,6,8,9 Neuraminidase A

	Store at (°C)	Concentration
Glycoprotein Denaturing Buffer	-20	10 X
NP-40	-20	10 %
GlycoBuffer 2	-20	10 X

O-Glycosidase

Featured Video

Overview of Glycobiology

Substrate Specificity:

Product Source

Reagents Supplied

Application Features

Unit Definition

Reaction Conditions

Storage Temperature

Storage Conditions

Heat Inactivation

Molecular Weight

Companion Products

Product Citation Tool

Featured Videos

Overview of Glycobiology

Other Products You May Be Interested In

O-Glycosidase & Neuraminidase Bundle

Protein Deglycosylation Mix II

α2-3,6,8,9 Neuraminidase A

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