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Description
O-GlycoproteaseProduct Specification Synonyms O glycosidase Purity 95% by SDS PAGE Storage Buffer 20 mM Tris HCl100 mM NaCl pH 7. 5 Stability & Storage This product should be stored at 20 and can be stored for at least 12 months. Avoid repeated freezing and thawing. Background O Glycoproteinase is expressed in E. coli and is a highly specific glycoproteinase that can specifically recognize serine or threonine residues of mucin type O linked sugars (whether
Product Specification
| Synonyms | O-glycosidase |
| Purity | ≥95% by SDS-PAGE |
| Storage Buffer |
20 mM Tris-HCl，100 mM NaCl ，pH 7.5 |
| Stability & Storage |
This product should be stored at -20℃ and can be stored for at least 12 months. Avoid repeated freezing and thawing. |
Background
O-Glycoproteinase is expressed in E. coli and is a highly specific glycoproteinase that can specifically recognize serine or threonine residues of mucin-type O-linked sugars (whether sialylated or not) and cleave them at their N-termini. It can produce O-glycopeptides suitable for mass spectrometry analysis. The enzyme contains a 6X His-Tag, which can be easily removed after the reaction.
Components
|
Components |
Size 1 |
Size 2 |
|
O-glycoprotease |
20U |
100U |
|
1M Tris-HCl（pH=8.0） |
1.0ml |
1.0ml×5 |
|
1M DTT |
0.1ml |
0.2ml |
|
1% SDS |
0.2ml |
1.0ml |
|
500mM iodoacetic acid |
0.1ml |
0.5ml |
Protocol
Instructions (for reference only)
- Mix 10µg of glycoprotein with 20 mM Tris-HCl pH 8.0, 40 mM DTT, 0.1% SDS solution and make the total volume 50µl.
- Heat at 95°C for 10 minutes.
- Cool to room temperature.
- Add 3µl 500mM iodoacetic acid solution.
- Leave at room temperature for 30 minutes.
- Transfer the solution to an ultrafiltration centrifuge tube.
- Centrifuge at 12000 x g for 4 minutes and discard the flow-through.
- Add 400µl 20mM Tris-HCl, pH 8.0 solution.
- Centrifuge at 12000 x g for 4 minutes and discard the flow-through. Add 400µl 20mM Tris-HCl, pH 8.0 solution and repeat once.
- Add 20µl 20mM Tris-HCl, pH 8.0 solution.
- Add 1µl O-glycoproteinase and mix gently.
- Incubate at 37˚C for 2-18 hours.
- Centrifuge at 12000 x g for 4 minutes and collect the supernatant.
Unit Definition
Shipping Notes
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Exchange/Return Notes
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