Endo F3

Product Description 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. Molecular weight 30,000 daltons 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 Defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of porcine fibrininogen in 1 minute at 37°C, pH 4.5. Cleavage is monitored by SDS-PAGE. Formulation The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, pH 7.5 Specificity Endo F3 cleaves free or Asparagine-linked triantennary or fucosylated biantennary oligosaccharides,as well as triamnnosyl chitobiose core structures. 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. The recombinant version is not glycosylated, which may result in properties differing from the native protein. Quality & Purity Endo F3 is tested for contaminating protease as follows: 10 µg of denatured BSA is incubated at 37°C for 24 hours 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 Several days exposure to ambient temperatures will not reduce activity. Stable at least 12 months when stored properly. Directions for use 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 F3 to the reaction. Incubate 1 hour at 37°C. Monitor cleavage by SDS-PAGE. The production host strain has been extensively tested and does not produce any detectable glycosidases.

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