Endo F3 cleaves free or Asparagine-linked triantennary or α(1-6) fucosylated biantennary oligosaccharides,as well as triamnnosyl chitobiose core structures. Nonfucosylated biantennary glycans will also be cleaved, 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. α(1-3) fucosylation will inhibit enzymatic activity. There is no activity on oligomannose and hybrid molecules.

Additional Endo F Products
Endoglycosidase F1 releases bianatennary and some hybrid glycans
Endoglycosidase F2 will release high mannose and biantennary N-glycans
Source: Recombinant Elizabethkingia miricola in E. Coli

EC: 3.2.1.96

Alternative Names: Endo F3, Endoglycosidase F3, endo-β-N-acetylglucosaminidase F

Specificity:
Cleaves all asparagine-linked triantennary or α(1-6) fucosylated biantennary oligosaccharides, as well as triamannosyl chitobiose core structures. α(1-3) fucosylation will inhibit enzymatic activity. Nonfucosylated biantennary glycans will also be cleaved, but at a 40x reduced rate. (Note: The recombinant version is not glycosylated, which may result in properties differing from the native protein.)

Contents:
Ludger Endoglycosidase F3 - Kit contents
60 µL aliquot of enzyme (0.3 U) in 20 mM Tris-HCl, pH 7.5
5x Reaction Buffer - 250 mM sodium acetate, pH 4.5

Specific Activity: >25 U/mg
Activity: >5 U/mL

Molecular weight: 30,000 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
4. Add 2.0 µL of Endo F3. 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 porcine fibrinogen in 1 minute at 37°C, pH 4.5. Cleavage is monitored by SDS-PAGE (cleaved fibrinogen 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 F3 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.