- Determination of protein and/or peptide glycosylation level
- Analysis of N-glycan functional roles
- Reduction of protein heterogeneity
- MALDI-TOF or other mass spectrometry
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
Recombinant Escherichia coli encoding PNGase F.
Specific cleavage of N-glycan (GluNAc-Asn) bonds in glycoproteins and glycopeptides.
PNGase F (Glycopeptidase F) will not cleave N-glycan groups 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℃ at pH 8.6.
- Molecular weight : approx. 36.5 kDa (SDS-PAGE)
(Note) Depending on proteolytic activity of host bacteria, recombinant PNGase F may have 15 or 18 additional animo acid residues attached to its N-terminus. However, PNGase F glycosylase activity has been confirmed to be identical to that of native PNGase F via experiments using RNase B and Fetuin as substrates.
- Optimum pH: 8.6