GlycOCATCH™
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GlycOCATCH™ Affinity Purification
Enrichment resin for affinity purification of mucin-type O-glycoproteins and peptides
GlycOCATCH is an affinity resin for purification of mucin-type 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.
GlycOCATCH Affinity Purification is provided in a convenient spin column format. The product contains:
- 4 GlycOCATCH Affinity Purification Microspin columns, each with a capacity to bind 50 µg mucin-type O-glycoprotein (200 µg in total)
- SialEXO Lyophilized 200 units, for removal of sialic acids
- OpeRATOR Lyophilized 200 units, for an optional elution method of the bound O-glycoprotein
Why GlycOCATCH™ Affinity Purification?
- Targeted O-glycoprotein enrichment
- Specific clean-up of complex samples
- Sample preparation for improved MS analysis
Available Products
GlycoCATCH™ Affinity Purification Microspin 0.2 mg
Enrichment resin for affinity purification of 0.2 mg mucin-type O-glycoproteins and peptides
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Product Information
Unit Defintion
- 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.
Contents
GlycOCATCH Affinity Purification 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 Lyophilized (200 units) supplied in TBS pH 7.6, with no preservatives added.
- 1 vial of OpeRATOR Lyophilized (200 units) supplied in TBS pH 7.6, with no preservatives added.
Content and Storage
GlycOCATCH Affinity Purification is shipped cold. Content should be stored at different temperatures upon arrival!
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.
Resources
Instruction for Use
Product Specifications
Safety Data Sheet
Certificate of Analysis
Find your certificate of analysis »SmartEnzymes™ for O-glycosylation Analysis using Mass Spectrometry
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Applications
Selective enrichment of O-glycoproteins from human serum for improved LC-MS/MS detection and glycosylation-focused proteomic analysis.
Selective enrichment of IgA O-glycopeptides for improved detection and characterization of hinge-region glycosylation.
FAQ and Support
When and why should I use the OpeRATOR elution procedure?
The OpeRATOR enzyme digests O-glycosylated proteins N-terminally to the serine (S) and threonine (T) glycosylation sites. OpeRATOR therefore facilitates mapping of O-glycosylation sites by LC-MS and as the digested products no longer bind to the GlycOCATCH resin, elution and sample preparation can be achieved in a single step. However, this requires the O-glycosylation sites to be spaced closely together for the OpeRATOR digestion to yield peptides of a size suitable for analysis by LC-MS/MS.
Is it possible to digest the glycoprotein with OpeRATOR first and then enrich the O-glycopeptides on GlycOCATCH?
No, when the glycoprotein is digested with OpeRATOR, the binding capacity of GlycOCATCH is lost.
Can I load a digested protein solution directly onto the GlycOCATCH column?
Yes. IMPORTANT! Before you load your sample make sure that the protease in solution is inactivated and that the pH, buffer, chaotrope and NaCl concentrations are within the recommended ranges according to the instructions.
Does GlycOCATCH bind all O-glycosylated proteins and peptides?
No, GlycOCATCH is designed to bind specifically to mucin type (core 1) O-glycosylated proteins and peptides. The performance is significantly enhanced by removal of sialic acids using SialEXO.
Legal & Disclaimers
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Genovis products are intended for research use only. They are not intended to be used for therapeutic or diagnostic purposes in humans or animals. All goods and services are sold subject to Genovis’ General Terms and Conditions of Sale.
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