- Assay a protein for the presence of N-Glycan
- Examine the functional role N-Glycan plays in a protein's activity
- Remove N-Glycan from a protein sample to facilitate glycan moiety analysis or reduce peptide heterogeneity prior to proteomic analysis
Denature buffer : 1% SDS / 1M Tris-HCl (pH8.6) 500 μl
Native buffer : 1M Tris-HCl (pH 8.6) 500 μl
Stabilized solution : 5% Nonidet P-40 500 μl
Control Glycoprotein : 10 mg/ml Bovine Fetuin 10 μl
Escherichia coli carrying the plasmid containing the gene encoding PNGase F.
Specific cleavage of N-glycans (GluNAc-Asn bonds) in glycopeptidase or glycoproteins.
PNGase F (Glycopeptidase F) is not able to cleave N-glycans from glycoproteins when the innermost GlcNAc residue is linked to an alpha1-3 Fucose residue. This modification is found in plant glycoproteins as well as some insect glycoproteins.
Definition of activity
One unit is the amount of enzyme required to hydrolyze 1 μmol of dansyl fetuin glycopeptide within 1 minitue at 37℃, pH8.6.
- Molecular weight : approx. 36,500 (SDS-PAGE)
(Note) Depending on proteolytic activity of host bacteria, the recombinant GPF may have 15 or 18 additional animo acid residues at its amino terminus. However, its activity on glycoproteins was confirmed to be identical to that of native GPF, by the experiments using RNase B and Fetuin as its substrates.
- Optimum pH: pH8.6