Description
ImpaRATOR is an O-glycan-dependent protease that digests proteins carrying mucin-type O-glycans, including sialylated species, N-terminally of glycosylated Ser and Thr residues.
ImpaRATOR Lyophilized
€1,215.00
Lyophilized enzyme for digestion of mucin-type O-glycoproteins and peptides, including sialylated O-glycan species
Detailed Information
| Application | |
|---|---|
| Product Group | |
| Source/Origin | Derived from P. aeruginosa, Expressed in E. coli (His-tagged), 97 kDa |
| Product Composition | |
| Digestion Site | |
| Specificity | Digests the protein backbone N-terminally of O-glycosylated Ser and Thr residues |
| Substrates | Native proteins with mucin-type O-glycosylation, including sialylated species |
| Reaction Conditions | 2 h (complex substrates may require longer incubation times) |
| Digestion Buffers | |
| Buffer Compatibility | Other buffers or pH (5.5-8.5 ) can be used but the reaction conditions need to be optimized |
| Inhibitors | |
| Unit Definition | One unit ImpaRATOR Lyophilized digests ≥ 95% of 30 µg etanercept at least at one site when incubated in TBS (50 mM Tris-HCl, 150 mM NaCl), pH 7.6 at 37°C for 30 minutes, For O-glycopeptide mapping applications, one unit ImpaRATOR Lyophilized digests 1 µg O-glycosylated protein when incubated in TBS, pH 7.6 at 37°C for 2 hours. |
FAQs
ImpaRATOR is a metalloprotease and thereby sensitive to chelating agents such as EDTA. Concentrations > 1 mM EDTA results in complete inhibition of the enzyme. In addition, the ImpaRATOR activity is inhibited by reducing agents and detergents.
Insufficient digestion during non-denaturing conditions can be caused by O-glycosylation sites within the glycoprotein inaccessible to the enzyme. In such cases, we recommend trying the following workflow: reduction, denaturation, carboxymethylation, buffer-exchange, and then digestion with ImpaRATOR. If detergent is added, make sure to include a detergent removal step prior ImpaRATOR digestion.
ImpaRATOR has a broad activity towards different O-glycan structures; however, the enzyme has limited activity towards sites with two adjacent O-glycosylated Ser/Thr residues. For complete information about O-glycan sites in glycoprotein substrates containing several sites with two adjacent O-glycosylated Ser/Thr residues, OpeRATOR may be a better option.
