>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<1.0 EU per 1 μg of the protein by the LAL method.
Measured by its ability to deglycosylate ribonuclease B under denatured conditions. >50% ribonuclease B (10 μg) is deglycosylated by 2.5 ng rFmPNGase F within 30 minutes, as measured under the described conditions.
Assay Protocol on www.RnDSystems.com.
E. coli-derived Ala41-Asn354 with N-terminal Met and 6-His tag
Recombinant F. meningosepticum PNGase F (Catalog # 9109-GH) from R&D Systems and a leading competitor are able to deglycosylate 10 μg of RNase B at 37 °C in one hour. The E. coli-produced enzyme from R&D systems offers a better value than the competition.
Background: PNGase F
PNGase F, peptide N-glycosidase F from Flavobacterium meningosepticum, catalyzes the hydrolysis of asparagine-linked high mannose, as well as hybrid and complex oligosaccharides from glycoproteins (1). Unlike glycosidases that hydrolyze glycosidic bonds, PNGase F is an amidase that cleaves the beta-aspartylglucosamine bond between the innermost GlcNAc of N-glycans and asparagine residues of glycoproteins (2). The enzyme is highly active on various N-glycans except those with the innermost GlcNAc modified with alpha 1-3-linked core fucose, which is commonly found on plant glycoproteins (3). Cleavage with PNGase F will convert the asparagine residue to an aspartic residue, allowing identification of the glycosylation site by mass spectrometry (4). This purified enzyme is compatible with glycan analysis using mass spectrometry.
Elder, J.H. and Alexander, S. (1982) Proc. Natl. Acad. Sci. USA 79:4540.
Maley, F. et al. (1989) Anal. Biochem. 180:195.
Tarentino, A.L. and Plummer, T.H. (1994) Methods Enzymol 230:44.
Zhang, H. et al. (2003) Nat. Biotechnol. 21:660.
Peptide-N4-(N-acetyl-beta-D-glucosaminyl)asparagine Amidase F
The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.