| Species : |
Streptomyces plicatus |
| Source : |
E. coli |
| Tag : |
Non |
| Description : |
Endo H cleaves Asparagine-linked hybrid or high mannose oligosaccharides, but not complex oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetyl chitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Detergent and heat denaturation may increase the rate of cleavage for some glycoproteins.
|
| Form : |
Sterile-filtered solution |
| EC : |
3.2.1.96 |
| Specificity : |
Endo H cleaves Asparagine-linked hybrid or high mannose oligosaccharides, but not complex oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Detergent and heat denaturation may increase the rate of cleavage for some glycoproteins. |
| Contents : |
60 μL aliquot of enzyme (300 mU) in 20 mM Tris-HCl, 25 mM NaCl, 1 mM EDTA (pH 7.5).
200 μL 5× Reaction Buffer 5.5 (250 mM sodium phosphate, pH 5.5)
Denaturation Solution-2% SDS, 1 M Beta-mercaptoethanol |
| Bio-activity : |
Specific Activity: > 40 U/mg
Activity: > 5 U/mL |
| Molecular Mass : |
29 kDa |
| Suggested usage : |
1. Add up to 200 μg of glycoprotein to Eppendorf tube. Adjust to 37.5 μL final volume with deionized water.
2. Add 10 μL 5× Endo H Buffer and 2.5 μL of Denaturation Solution (SDS/-ME). Heat at 100 centigrade for 5 minutes.
3. Cool, and then add 2.0 μL of Endo H to the reaction. Incubate 3 hours at 37 centigrade. |
| Unit Definition : |
One unit of Endo H activity is defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 μmole of denatured Ribonuclease B. Cleavage is monitored by SDS-PAGE (cleaved Ribonuclease B migrates faster). |
| Purity : |
Endoglycosidase H 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.
The production host strain has been extensively tested and does not produce any detectable glycosidases. |
| Applications : |
Releases asparagine-linked hybrid or high mannose oligo saccharides, but not complex oligosaccharides. Endo H cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Detergent and heat denaturation may increase the rate of cleavage for some glycoproteins. |
| Stability : |
Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity. |
| Storage : |
Store enzyme at 4 centigrade. |
| Storage Buffer : |
The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, 25 mM NaCl, 1 mM EDTA (pH 7.5). |
