Releases unsubstituted, Ser/Thr-linked GalGalNAc from proteins; used with additional enzymes to remove more complex O-linked structures. Also known as Endo-O-Glycosidase™, Endo-α N-acetylgalactosaminidase, O-Glycosidase.
Recombinant gene from Streptococcus pneumoniae, expressed in E.coli.
The enzyme is highly specific, liberating Galβ1-3GalNAc from serine or threonine residues alone or as part of glycopeptides or glycoproteins. There is no apparent preference for serine over threonine. Substitution of the disaccharide core with sialic acid, fucose, N-acetylglucosamine or N-acetylgalactosamine residues, prevents cleavage of the Galβ1-3GalNAc-Ser/Thr linkage. Absence of the galactose residue or the acetamido group on the innermost N-acetylgalactosamine residue also prevents cleavage of the Galβ1-3GalNAc-Ser/Thr linkage.
O-Glycanase is included in our Enzymatic Deglycosylation Kit for N-Linked & Simple O-Linked Glycans (GK80110), which may be used in conjunction with our prO-LINK Extender™ Kit for Complex O-Linked Glycans (GK80115).
WS0059 5X reaction buffer A [250 mM sodium phosphate pH 5.0]
20 mM Tris HCl pH 7.5 containing 25 mM NaCl.
5X concentrated buffer which when diluted gives 50 mM sodium phosphate pH 5.0.
5.5 / 5.0 - 7.0
One unit is defined as the amount of enzyme required to catalyze the release of 1 μmole of Galβ1-3GalNAcα1-p-nitrophenol per minute at 37°C, pH 5.5.
Size: 50 mU (40 µl)
Concentration: ≥ 1.25 U/ml
Product Code: GK80090
Q. Can GK80090 O-Glycanase be used with detergents and reductants?
A. The enzyme can be used with sodium dodecyl sulfate (SDS) and β-mercaptoethanol (βME) at certain levels.
The 5x Denaturation Solution used in the GK80110 kit is 2% sodium dodecylsulfate (SDS) and 1 M βME, and 2.5μl is used in a reaction volume of around 50μl giving a final reaction concentration of 0.1% SDS, 50mM bME. Denaturation Solution and 5x Incubation Buffer are added to the glycoprotein substrate and the mixture is heated to 100°C for 5 minutes. After cooling, 2.5μl of a Detergent Solution (15% NP-40, final reaction concentration 0.75%) is added to counteract the inhibitory effect of SDS on PNGase F. This is followed by the addition of N-Glycanase, Sialidase A (GK80040) and O-Glycanase (GK80090), with incubation for 3 hours at 37°C.
Product Safety Documentation for GK80090:
|WS0059||5x Incubation Buffer A|