O-Glycosidase

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 40,000,000 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 40,000,000 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

Companion Products

• O-Glycosidase & Neuraminidase Bundle
• α2-3,6,8 Neuraminidase
• PNGase F
• PNGase F (Glycerol-free)
• Endo H_f
• Endo H

1. 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).
2. To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required.
3. 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.
4. Under denaturing conditions the enzyme activity is increased two-fold. This observation is substrate dependent.

1. Koutsioulis, D., Landry, D. and Guthrie, E.P. (2008). Glycobiology. 18, 799-805.
2. Morgan, W.T.J. and Elson, L.A. (1994). Biochem. J.. 28, 988-995.

1. O-Glycosidase Application Note 1 (P0733)
2. O-Glycosidase (P0733)
3. Endo-α-N-Acetylgalactosaminidase Application Note 1

• AppNote_Enzymatic_Deglycosylation_of_Protein_Containing_Core_1_O-glycans_with_O-Glycosidase
• Analysis of a Fusion Protein using the Protein Deglycosylation Mix II and Mass Spectrometry
• Enzymatic Deglycosylation of a Protein Containing Core 1 O-glycans with O-Glycosidase

• Endoglycosidase Selection Chart

1. What are the typical reaction conditions for O-Glycosidase?
2. I tried using O-Glycosidase on my glycoprotein and didn't see removal of the carbohydrate. What could be the problem?
3. How much O-Glycosidase should I use to remove my carbohydrate under native conditions?
4. Do detergents inhibit O-Glycosidase?
5. Can Neuraminidase be used together in a digest with PNGase F and O-Glycosidase?
6. Can I double digest PNGase F and O-Glycosidase?
7. What is the difference between PNGase F, Endo H and O-Glycosidase?
8. 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?
9. What are glycosidases and their uses?
10. Is it necessary to treat my glycoprotein concomitantly with Neuraminidase and O-Glycosidase?
11. How do I inhibit O-Glycosidase?

• Glycoproteomics Brochure
• Glycoproteomics Technical Guide

• Using Glycosidases to Remove Trim or Modify Glycans on Therapeutic Protein
• O-glycan analysis of therapeutic proteins enabled by O-glycoprotease

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]

• P0733S_L_v1

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
• P0733S_v1_10057983
• P0733S_v1_10062547
• P0733S_v1_10077195
• P0733L_v1_10082483
• P0733L_v1_10097236
• P0733S_v1_10084150
• P0733L_v1_10106703
• P0733S_v1_10112430
• P0733L_v1_10128264
• P0733S_v1_10126416
• P0733S_v1_10149065
• P0733L_v1_10149063

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

• O-Glycosidase
• GlycoBuffer 2
• Glycoprotein Denaturing Buffer
• NP-40

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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.

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.