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Endo F1 cleaves high mannose and some hybrid type N-glycans from peptides and proteins.
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.
Source: Recombinant Elizabethkingia miricola in E. Coli
Alternative Names: Endo F1, Endoglycosidase F1, endo-β-N-acetylglucosaminidase F
Endo F1 Specificity: Cleaves all asparagine-linked high mannose and some hybrid oligosaccharides
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
included with E-EF01 and E-EF01-20: 1 vial: 5x Reaction Buffer, 250 mM sodium phosphate, pH5.5
Activity ≥ 17 U/ml
Specific Activity ≥ 16 U/mg
Molecular Weight: 32 kD
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).
Formulation: The enzyme is provided as a sterile-filtered solution in 20mM Tris-HCl, pH 7.5
Storage: Store enzyme at 4°C. Do not freeze.
Stability: Several days exposure to ambient temperatures will not reduce activity. Stable at least 12 months when stored properly.
Ludger Endo F1 cleaves Asparagine-linked high mannose or hybrid oligosaccharides. 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.
Quality & Purity
Ludger Endo F1 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.
Directions for use
- Add up to 200 μg of glycoprotein to an Eppendorf tube.
- Adjust to 38 μl final volume with de-ionized water.Add 10 μl 5x Reaction Buffer 5.
- Add 2.0 μl of Endo F1 to the reaction. Incubate 1 hour or more at 37°C.
- Monitor cleavage by SDS-PAGE.