Description
PNGase F, Recombinant Glycosidase
PNGase F is a recombinant glycosidase cloned from Elizabethkingia miricola and overexpressed in E. coli . PNGase F has a molecular weight of 36kDa. PNGase F catalyzes the cleavage of N-linked oligosaccharides between the innermost GlcNAc and asparagine residues of high mannose, hybrid and complex oligosaccharides from N-linked glycoproteins (see figure). PNGase F will not remove oligosaccharides containing Alpha-(1,3)-linked core fucose commonly found on plant glycoproteins.
Unit Definition: One unit of PNGase F will catalyze the deglycosylation of 1 nanomole of denatured Ribonuclease B (RNase B) in one minute at 37°C. One Promega unit is equal to 1 IUB milliunit.
Molecular Weight: PNGase F has a molecular weight of approximately 36kDa.
Physical Form: PNGase F is supplied as a liquid in 20mM Tris-HCl (pH 7.5 at 25°C), 50mM NaCl and 5mM EDTA at a concentration of 10,000u/ml.
Characterizing whether the protein is glycosylated
Determining the location of glycosylation on the protein
Characterizing the glycan structure
Protein trafficking
Twenty micrograms of various denatured glycoprotein standards were treated with 10 units of Promega recombinant or native PNGase F for 1 hour at 37°C. Gel shifts indicating deglycosylation were monitored by SDS-PAGE.
