Endo F2 cleaves N-linked (asparagine-linked) biantennary oligosaccharides from glycoproteins. It also will cleave high mannose glycans but at a 40x reduced rate. 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.

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.

Additional Endo F Products:
Endoglycosidase F1 releases bianatennary and some hybrid glycans
Endoglycosidase F3 will release triantennarry and fucosylated biantennary N-glycans
Source: Recombinant Elizabethkingia miricola in E. Coli

EC: 3.2.1.96

Alternative Names: Endo F2, Endoglycosidase F2, endo-β-N-acetylglucosaminidase F2

Specificity:
Cleaves all asparagine-linked biantennary oligosaccharides and high mannose (though at a 40X reduced rate) N-glycans from peptides and proteins

Contents:
Ludger Endoglycosidase F2 - Kit contents
60 µL aliquot of enzyme (0.3 U) in 10 mM sodium acetate 25mM NaCl, pH 4.5
5x Reaction Buffer - 250 mM sodium acetate, pH 4.5

Specific Activity: >20 U/mg
Activity: >5 U/mL

Molecular weight: 32,000 daltons

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 5x Reaction Buffer 4.5
3. Add 2.0 µL of Endo F2. Incubate 1 hour at 37°C.

Specific Activity:
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°C, pH 5.5. Cleavage is monitored by SDS-PAGE (cleaved finbrinogen migrates faster).

Storage: Store enzyme at 4°C.

Stability: Stable at least 12 months when stored properly. Several days exposure to ambient tempertures will not reduce activity.

Purity: Endoglycosidase F2 is tested for contaminating protease as follows; 10 µg of denatured BSA is incubated for 24 hours at 37°C 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.