Endo F1 cleaves Asparagine-linked high mannose and some hybrid oligosaccharides. Core fucosylation reduces the activity by 50 fold. Endoglycosidase F1 will hydrolyze sulfate containing high-mannose chains. 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.

Additional Endo F Products:
Endoglycosidase F2 releases biantennary and high mannose glycans (at a 40X reduced rate)
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 F1, Endoglycosidase F1, endo-β-N-acetylglucosaminidase F

Endo F1 Specificity: Cleaves all asparagine-linked high mannose and some hybrid oligosaccharides

Contents:
Ludger Endoglycosidase F1 - Kit contents
60 µL aliquot of enzyme (1 U) in 20 mM Tris-HCl, pH 7.5
5x Reaction Buffer – 250 mM sodium phosphate, pH 5.5

Specific Activity: >16 U/mg
Activity: >17 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 5.5
3. Add 2.0 µL of Endo F1. 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 Ribonuclease B (RNase B) in 1 minute at 37°C, pH 5.5. Cleavage is monitored by SDS-PAGE (cleaved RNase B 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 F1 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.