| Species : |
Streptococcus pneumoniae |
| Source : |
E. coli |
| Tag : |
Non |
| Description : |
O-glycosidase cleaves only unsubstituted Gal-β(1-3)GalNAc-α disaccharides attached to the serine or threonine residues of glycoproteins or glycopeptides.
Substitutions such as sialic acid, galactose, fucose or N-acetylglucosamine must first be removed with the appropriate exoglycosidase before treatment with Endo-O-Glycosidase. At minimum, a sialadase such as Sialadase Au α(2-3,6,8,9) is almost always required to remove sialic acids.
Kit includes enzyme plus reaction buffer. Sufficient for up to 60 reactions. |
| Form : |
Sterile-filtered solution |
| EC : |
3.2.1.97 |
| Specificity : |
O-glycosidase leaves only unsubstituted Gal-β(1-3)GalNAc-α disaccharides attached to the serine or threonine residues of glycoproteins or glycopeptides.
Substitutions such as sialic acid, galactose, fucose or N-acetylglucosamine must first be removed with the appropriate exoglycosidase prior to treatment with Endo-O-Glycosidase. At minimum, a sialadase such as Sialadase Au α(2-3,6,8,9), is almost always required to remove sialic acids. |
| Contents : |
60 μL aliquot of enzyme (75 mU) in 50 mM sodium phosphate, pH 7.5.
5× Reaction Buffer 5.0 (250 mM sodium phosphate, pH 5.0) |
| Bio-activity : |
Activity: > 1.25 U/mL
Specific Activity: > 12 U/mg
Defined as the amount of enzyme required to produce 1 μmole of p-nitrophenol (pNP) in 1 minute at 37 centigrade, pH 5.0 from p-nitrophenyl-2-acetamido-2-deoxy-3-O-(β-D-galactopyranosyl)-α-D-galactopyranoside. |
| Molecular Mass : |
180 kDa |
| Suggested usage : |
1. Add up to 100 μg of glycoprotein to a tube.
2. Add de-ionized water to a total of 13 μL.
3. Add 4 μL of 5× Reaction Buffer 5.0.
4. Add 1 μL of Sialadase Au α(2-3,6,8,9)
5. Add 2 μL of O-Glycosidase.
6. Incubate for 1 hour at 37 centigrade. |
| Unit Definition : |
One unit of O-Glycosidase is defined as the amount of enzyme required to produce 1 μmole of p-nitrophenol (pNP) in 1 minute at 37 centigrade, pH 5.0 from p-nitrophenyl 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl) alpha-D-galactopyranoside. |
| Purity : |
O-Glycosidase is tested for contaminating protease as follows; 10 μg of denatured BSA is incubated for 24 hours at 37 centigrade with 2 μL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation. |
| Stability : |
Stable at least 12 months when stored properly. Several days exposure to ambient temperature will not reduce activity. Active for at lease 5 days under reaction conditions. |
| Storage : |
Store enzyme at 4 centigrade. |
| Storage Buffer : |
The enzyme is provided as a sterile-filtered solution in 50 mM Sodium phosphate pH 7.5. |
