OpeRATOR™ Lyophilized

Lyophilized enzyme for digestion of mucin-type O-glycoproteins and peptides

Available Products

  • Lyophilized

OpeRATOR is an O-glycan-specific protease that digests proteins carrying core 1 O-glycans, N-terminally of the glycosylated (core 1) Ser and Thr residues.

OpeRATOR Lyophilized is available as a lyophilized powder in 2000 unit vials for digestion of 2 mg O-glycosylated protein. Removal of sialic aids is required for optimal performance, and 2000 units if SialEXO Lyophilized is therefore included in the purchase.

Why OpeRATOR™ 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

  • OpeRATOR Lyophilized 2000 units

    Lyophilized enzyme for digestion of 2 mg mucin-type O-glycoproteins and peptides

    G2-OP1-020

    €1,251.00

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

    Product Information

    Unit Defintion

    One unit digests ≥ 90% 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.

    Contents

    OpeRATOR Lyophilized includes:

    • 1 vial of OpeRATOR Lyophilized (G1-OP1-020) supplied lyophilized in TBS pH 7.6, with no preservatives added.
    • 1 vial of SialEXO Lyophilized (G1-SM1-020) supplied lyophilized in TBS pH 7.6, with no preservatives added.

    Storage

    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.

    Resources

    Applications

    Citations

    FAQ and Support

    If digestion is insufficient, it could be caused by the O-glycans having a sterically inaccessible location on the sample that the enzyme cannot reach. In such cases, we recommend trying the following workflow: reduction, denaturation, carboxymethylation, rebuffering, and then digestion with SialEXO and OpeRATOR.

    The relative abundance and site occupancy of O-glycosylation may vary a lot in a glycoprotein sample, and this will result in the generation of peptides with variable sizes. In some cases, the peptides contain more than one O-glycan, i.e. OpeRATOR does not digest at every site in every molecule. However, the missed digestion sites differ from molecule to molecule, and the compiled data will provide valuable information on the O-glycosylated sites.

    OpeRATOR is a metalloprotease and as such highly sensitive to chelating agents such as EDTA. Concentrations >5mM lead to complete inhibition of the enzyme. OpeRATOR activity is also moderately inhibited by ZnCl2.

    The activity of OpeRATOR is significantly decreased in the presence of sialic acids. We recommend using SialEXO to remove the sialic acids. If this is not an option, then we suggest trying to digest both with and without SialEXO to evaluate the impact for your specific samples and requirements.

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

    Find all FAQ here

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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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