SmartEnzymes SialEXO workflow

Hydrolysis of Sialic Acids

The SialEXO products are sialidases for the removal and analysis of sialic acids. The enzymes are active on both N- and O-linked glycans present on native glycoproteins or released glycan structures.

SialEXO is used to pretreat O-glycosylated proteins prior to digestion with OpeRATOR or OglyZOR. Other applications include reduction of sample complexity, charge variant analyses and exoglycosidase arrays.
 

  • Efficient desialylation of N-linked, O-linked and released glycans
  • Fast removal of sialic acids for enhanced analysis of protein charge variants
  • Available immobilized in ready-to-use spin columns
Sialic acids on N- and O-linked glycans
30 min - 2 h reaction
No need for co-factors
α2-3, α2-6 and α2-8-linked sialic acids

SialEXO® Lyophilized

Lyophilized enzyme mix for desialylation of glycoproteins

SialEXO® 2-3 Lyophilized

Lyophilized enzyme for hydrolysis of ɑ2-3-linked sialic acids

SialEXO® Immobilized

Immobilized enzymes for desialylation of glycoproteins in spin columns

About SialEXO®

SialEXO workflow

The SialEXO products are sialidases for efficient desialylation of N- and O-glycosylated proteins.

SialEXO Lyophilized consists of two sialidases, each having unique activities and they work simultaneously. One enzyme has a broad activity for α2-3, α2-6 and α2-8-linked sialic acids, and the other one (SialEXO 2-3) quickly hydrolyzes sialic acids with α2-3-linkage. SialEXO Lyophilized removes all sialic acids on native proteins within 2 h.

SialEXO hydrolyzes glycoproteins under native conditions and displays activity in pH ranging from 6.5 to 9.

The enzymes are derived from Akkermansia muciniphila and expressed in E. coli. Both enzymes contain a His-tag and the molecular weights of the enzymes are 43 kDa and 66 kDa.

SialEXO® Lyophilized

Lyophilized enzyme mix for desialylation of glycoprotein.

SialEXO Lyophilized is available as a lyophilized powder in 2000 unit vials for desialylation of 2 mg glycoprotein.
 

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

Available Products

SialEXO Lyophilized 2000 unitsLyophilized enzyme mix for desialylation of 2 mg glycoproteinG1-SM1-020736.00Buy / 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 of SialEXO Lyophilized hydrolyzes sialic acids from ≥ 90% of 1 µg glycoprotein (fetuin) when incubated in 20 mM Tris pH 6.8 at 37 °C for 2 h.

Content and Storage

SialEXO Lyophilized is reconstituted by addition of water. No preservatives added. After reconstitution, the enzyme is stable for 1 month at +4-8°C. The product is shipped at ambient temperature, and should be stored at -20°C upon arrival.

Instructions

Product Specification

Safety Data Sheet

Certificate of Analysis

SialEXO® 2-3 Lyophilized

Lyophilized enzyme for hydrolysis of ɑ2-3-linked sialic acids.

SialEXO 2-3 Lyophilized is available as a lyophilized powder in 500 unit vials for desialylation of 0.5 mg glycoprotein.

In contrast to SialEXO, which is a sialidase mix, SialEXO 2-3 is an α2-3 specific sialidase, allowing targeted analysis of α2-3 linked sialic acids only. Efficient α2-3 desialylation of O- and N-glycosylated proteins can be achieved within 1 h.

SialEXO 2-3 hydrolyzes glycoproteins under native conditions and displays activity in pH ranging from 7.0 to 9.0.

The enzyme is derived from Akkermansia muciniphila and expressed in E. coli. The enzyme has a His-tag and the molecular weight is 66 kDa.
 

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

Available Products

SialEXO 2-3 Lyophilized 500 unitsLyophilized enzyme for hydrolysis of ɑ2-3-linked sialic acids from 0.5 mg glycoproteinG1-SD2-005607.00Buy / 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 of SialEXO 2-3 Lyophilized hydrolyzes α2-3 linked sialic acids from ≥ 90% of 1 µg glycoprotein (fetuin) when incubated in 20 mM Tris pH 7.5 at 37 °C for 1 h.

Content and Storage

SialEXO 2-3 Lyophilized is reconstituted by addition of water. No preservatives added. After reconstitution, the enzyme is stable for 1 month at +4-8°C. The product is shipped at ambient temperature, and should be stored at -20°C upon arrival.

Product Specification

Safety Data Sheet

Certificate of Analysis

SialEXO® Immobilized

Immobilized enzymes for desialylation of glycoproteins in spin columns.

The SialEXO Immobilized spin columns contain SialEXO enzyme covalently coupled to agarose beads, for desialylation of glycoproteins without contaminating the final preparation with enzyme. It hydrolyzes sialic acids (α2-3, α2-6 and α2-8) on native glycoproteins and released glycans.

The Microspin columns are available for desialylation of 2 x 0.5 mg, 5 x 0.5 mg, or 10 x 0.5 mg glycoprotein.
 

  • Easy-to-use spin columns
  • Increased enzyme-to-substrate ratio for improved digestion efficiency
  • No enzyme in the final sample

Available Products

SialEXO Immobilized Microspin 2 x 0.5 mgImmobilized enzymes for desialylation of 2 x 0.5 mg glycoprotein in spin columnsG1-SM6-010491.00Buy / Request a Quote
SialEXO Immobilized Microspin 5 x 0.5 mgImmobilized enzymes for desialylation of 5 x 0.5 mg glycoprotein in spin columnsG1-SM6-0251,040.00Buy / Request a Quote
SialEXO Immobilized Microspin 10 x 0.5 mgImmobilized enzymes for desialylation of 10 x 0.5 mg glycoprotein in spin columnsG1-SM6-0501,730.00Buy / Request a Quote

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

Unit Definition

0.1 ml of settled SialEXO Immobilized agarose resin hydrolyzes sialic acids from > 90% of 0.5 mg C1 inhibitor in 30 min at room temperature in PBS pH 7.4 as monitored by ion exchange chromatography.

One Microspin column contains sufficient SialEXO-coupled agarose beads to desialylate 0.5 mg glycoprotein.

Content and Storage

SialEXO Immobilized is supplied in 20% ethanol, shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the product!

Instructions

Product Specification

Safety Data Sheet

Certificate of Analysis

FAQ and Support

Popular FAQ

Yes, SialEXO can be incubated together with OpeRATOR and/or OglyZOR in 20 mM Tris pH 6.8.

Yes, SialEXO contains a 6x His-tag.

Yes, you can. All three enzymes perform well in the same reaction buffer (20 mM Tris pH 6.8). There is no need to increase incubation time when combining the enzymes.

Yes, SialEXO displays activity on sialic acids with α2-3, α2-6 and α2-8 linked bonds of both N- and O-glycans.

No, SialEXO works both on glycoproteins in native conditions but can also be used on denatured proteins after buffer exchange.

 

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