Product Description Endo F3, Endoglycosidase F3, endo-beta-N-acetylglucosaminidase F Endo F3 cleaves free or Asparagine-linked triantennary or alpha-(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. Alpha 1-3 fucosylation will inhibit enzymatc activity. There is no activity on oligomannose and hybrid molecules.
Source recombinant Elizabethkingia meningosepticum (was Chryseobacterium meningosepticum)
EC 3.2.1.96 Specific Activity >25 U/mg Activity >5 U/ml Molecular weight 30,000 daltons
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).
Specifictity Cleaves all asparagine-linked triantennary or alpha-(1-6)-fucosylated biantennary oligosaccharides, as well as triamannosyl chitobiose core structures. Alpha 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 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
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.
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 37oC 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. | 
