| Species : |
Elizabethkingia miricola |
| Source : |
E. coli |
| Tag : |
Non |
| Description : |
Endo F2 cleaves N-linked (asparagine-linked) biantennary oligosaccharides from glycoproteins. It also will cleave high mannose glycans but at a 40× reduced rate. 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.
Endoglycosidase F2 is less sensitive to protein conformation than PNGase F and is, therefore, more suitable for deglycosylation of native proteins. However, for optimal results, denaturation of the glycoprotein is recommended. |
| Form : |
Sterile-filtered solution |
| EC : |
3.2.1.96 |
| Specificity : |
Cleaves all asparagine-linked biantennary oligosaccharides and high mannose (though at a 40× reduced rate) N-glycans from peptides and proteins |
| Contents : |
60 μL aliquot of enzyme (0.3 U) in 10 mM sodium acetate 25 mM NaCl, pH 4.5
5× Reaction Buffer-250 mM sodium acetate, pH 4.5 |
| Bio-activity : |
Activity: > 5 U/mL
Specific Activity: > 20 U/mg
Defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of denatured porcine fibrinogen in 1 minute at 37 centigrade, pH 5.5. Cleavage is monitored by SDS-PAGE (cleaved finbrinogen migrates faster). |
| Molecular Mass : |
32 kDa |
| Suggested usage : |
1. Add up to 200 μg of glycoprotein to an Eppendorf tube. Adjust to 38 μL final volume with de-ionized water.
2. Add 10 μL 5× Reaction Buffer 4.5
3. Add 2.0 μL of Endo F2. Incubate 1 hour at 37 centigrade. |
| Purity : |
Endoglycosidase F2 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. |
| 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. Do not freeze. |
| Storage Buffer : |
The enzyme is provided as a sterile-filtered solution in 10 mM sodium acetate, 25 mM NaCl, pH 4.5 |
