SmartEnzymes OglyZOR workflow

Hydrolysis of Core 1 O-glycans

OglyZOR is an endoglycosidase (O-Glycosidase) that specifically hydrolyzes core 1 O-glycans on native glycoproteins.

OglyZOR is used for removal of O-glycans for glycan analysis, confirmation of O-glycan presence and reduction of sample heterogeneity.
 

  • Highly efficient O-glycosidase
  • Reduces sample heterogeneity to unmask potentially critical protein modifications
  • Complete and robust removal of O-glycan core 1 disaccharides
O-glycans on glycoproteins
2-4h reaction
Requires removal of terminal sialic acids using SialEXO® (included)
Hydrolyzes core 1 type O-glycans

OglyZOR® Lyophilized

Lyophilized enzyme for hydrolysis of core 1 O-glycans on glycoproteins

About OglyZOR®

OglyZOR workflow

OglyZOR is an endoglycosidase (endo-α-N-Acetylgalactosaminidase) that catalyzes the hydrolysis of core 1, and to a limited extent core 3, O-glycans from native glycoproteins.

Removal of sialic acids is required for OglyZOR enzyme activity, and therefore, SialEXO Lyophilized composed of a mix of sialidases for removal of α2-3, α2-6 and α2-8-linked sialic acids, is supplied together with OglyZOR. Optimal activity is obtained at pH 6.5 to 7.5 and at 37°C.

OglyZOR is derived from Streptococcus oralis and expressed in E. coli. The enzyme contains a His-tag and the molecular weight is 227 kDa. SialEXO is derived from Akkermansia muciniphila and expressed in E. coli. The SialEXO enzymes contain His-tags and the molecular weights of the components are 42 kDa and 66 kDa, respectively.

OglyZOR® Lyophilized

Lyophilized enzyme for hydrolysis of core 1 O-glycans on glycoproteins.

OglyZOR Lyophilized is available as a lyophilized powder in 2000 unit vials for hydrolysis of core 1 O-glycans on 2 mg glycoprotein. It requires removal of sialic aids for activity, and is therefore provided together with 2000 units of SialEXO Lyophilized.
 

  • Flexible format for method development
  • Possible to combine enzymes for improved efficiency
  • Amenable to automated workflows
  • Ready to use – just add water!

Available Products

OglyZOR Lyophilized 2000 unitsLyophilized enzyme for hydrolysis of core 1 O-glycans on 2 mg glycoproteinG2-OG1-0201,017.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One unit digests ≥ 90% of O-glycans of 1 µg glycoprotein (TNFR) when incubated together with one unit of SialEXO in 20 mM Tris pH 6.8 at 37°C for 2 h.

Content and Storage

OglyZOR Lyophilized includes:

  • 1 vial of OglyZOR Lyophilized (G1-OG1-020) supplied in TBS pH 7.6, with no preservatives added.
  • 1 vial of SialEXO Lyophilized (G1-SM1-020) supplied in TBS pH 7.6, with no preservatives added.

The product is shipped at ambient temperature, and should be stored at -20°C upon arrival. After reconstitution, the enzymes are stable for 1 month at +4-8 °C.

Instructions

Product Specification

Safety Data Sheet

Certificate of Analysis

FAQ and Support

Popular FAQ

No, OglyZOR recognizes the core 1 disaccharide HexNAcHex (or GalNAcGal) and to some extend also the core 3 disaccharide HexNAc(2) (or GalNAcGlcNAc).

OglyZOR only works on desialylated O-glycans therefore is a sialidase (SialEXO) included with the product. SialEXO is not O-glycan specific and will therefore desialylate any N-glycans that are present.

Yes, we have performed successful experiments with OglyZOR, SialEXO and PNGaseF in the same reaction under native conditions O/N at pH ~7.0.  If denaturing reaction conditions are required for PNGaseF, the reaction with OglyZOR and SialEXO needs to be performed first under native conditions, then add PNGaseF and the required buffer.

Yes, this workflow enables MS/MS of O-glycosylated peptides with removed O-glycans.

 

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