Enzymes for O-glycans

Enzymes for O-glycans

GlycOCATCH™

GlycOCATCH is an enrichment resin for affinity purification of O-glycosylated proteins and peptides. 

The GlycOCATCH resin is provided in a convenient spin column format. The box contains 4 GlycOCATCH Microspin columns with a capacity to bind 50 µg O-glycoprotein each (200 µg total). GlycOCATCH is provided together with 200 units of SialEXO™, for removal of sialic acids, and 200 units of OpeRATOR™, an optional elution method of the bound O-glycoprotein.

 

Detailed Information

Product Description

GlycOCATCH is designed to specifically bind O-glycosylated proteins and peptides. The affinity resin is based on inactive OpeRATOR™ enzyme that has been engineered to bind O-glycosylated proteins and peptides with high affinity. O-glycoproteins and peptides are bound to the resin at pH 5-8.

GlycOCATCH is provided in a spin column format to allow easy-to-use enrichment of O-glycoproteins. The binding is performed in physiological buffers at pH 5-8 and combined with desialylation using SialEXO for 30 min. Due to the strong interaction between the GlycOCATCH resin and O-glycoproteins/peptides, the elution is performed with 8 M urea. Alternatively, the elution can be performed with the included, active OpeRATOR enzyme, that digests the bound O-glycoproteins N-terminally of the O-glycan sites. In this case, peptides are recovered under native conditions. Since asialylated O-glycans on O-glycoproteins are bound efficiently to the resin, the sialidase mix SialEXO™ is included in the box.

Binding Capacity and Unit Definitions

50 μl of the GlycOCATCH resin binds ≥ 50 μg asialylated etanercept when incubated in 10 mM sodium phosphate buffer pH 7.4, 150 mM NaCl at room temperature for 30 min.

One unit of SialEXO hydrolyzes sialic acids from ≥ 90% 1 µg glycoprotein (fetuin) when incubated in 20 mM Tris pH 6.8 at 37°C for 2h.

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

Content and Storage

The GlycOCATCH box (G3-OC6-002) includes:

  • 4 Microspin columns containing GlycOCATCH resin supplied in 20% EtOH, with no preservatives added. Each column includes sufficient material to bind 50 µg O-glycoprotein
  • 1 vial of SialEXO (200 units) supplied lyophilized in TBS pH 7.6, with no preservatives added.
  • 1 vial of OpeRATOR (200 units) supplied lyophilized in TBS pH 7.6, with no preservatives added.

The GlycOCATCH box is shipped cold. The GlycOCATCH resin should be stored at +4-8 °C upon arrival. Do not freeze the columns! The vials of SialEXO and OpeRATOR should be stored at –20 °C upon arrival. After reconstitution, SialEXO and OpeRATOR are stable for 1 month at +4-8 °C.

Applications

 

The GlycOCATCH Box

  • 4 x 50 ug GlycOCATCH Microspin columns 
  • 200 units SialEXO
  • 200 units OpeRATOR

Workflow Overview

  • Add SialEXO to the O-glycosylated protein or the complex sample
  • Transfer the sample to the GlycOCATCH column
  • Incubate for 30 min
  • Elute with 8 M urea, or with OpeRATOR

Key Characteristics

  • Enriches O-glycosylated proteins and peptides
  • Native binding, no requirement for binding buffers
  • 30 min - 2 h binding
  • Requires removal of sialic acids using SialEXO (included)
  • Elution is performed using 8 M urea or OpeRATOR (included)

How to order

For details on ordering please visit How to order. Click add to cart above to buy online.

Quotations

Please do not hesitate to contact us if you need a quotation or have inquiries on price of bulk quantities or licensing opportunities.

Documents

Instructions

 

 

Product specifications

 

 

Safety Data Sheet

 

 

FAQ and Support

The OpeRATOR enzyme digests O-glycosylated proteins N-terminally of the serine (S) and threonine (T) glycosylation sites. If you want to know at which S or T a specific O-glycan is sited, we recommend elution with OpeRATOR. We also recommend elution with OpeRATOR if you have a glycoprotein with suitable distances between the O-glycan sites and suitable distances between the O-glycans and the N/C-terminals, and you want to generate peptides of suitable size for your purpose.
No, when the glycoprotein is digested with OpeRATOR, the binding ability of GlycOCATCH is lost.
Before you load your sample, make sure that the protease in the solution is inactivated, and that the pH, buffer, chaotrope and NaCl concentrations are within the recommended ranges (according to the instructions).

Find all FAQ here

Patent and Disclaimer

GlycOCATCH™

All rights reserved. Aspects of GlycOCATCH™ technology are encompassed by pending patent applications in the name of Genovis AB.

The trademark GlycOCATCH™ is the property of Genovis AB.

For research use only. Not intended for any animal or human therapeutic or diagnostic use.

©2018 Genovis AB.