This product is a recombinant O-Glycosidase (endo-α-N-acetylgalactosaminidase, O-glycanase; glycosyl hydrolase family GH101; E.C. 3.2.1.97), cloned from Enterococcus faecalis and expressed in Escherichia coli with an N-terminal 8xHis tag. The 8xHis tag may be removed by digestion with FasTEV™ (Cat #GE0501), a TEV protease with enhanced stability and catalytic activity.
This enzyme catalyzes the hydrolysis of the unsubstituted O-glycans from Ser/Thr residues of glycoprotein substrates. It predominantly cleaves the Core 1 structure (T antigen).
Unit definition: One unit is defined as the amount of enzyme required to catalyze the release of 1 nmole of p-nitrophenol (pNP) from 500 μM p-nitrophenyl 2-acetamido-2-deoxy-3-O-(β-D-galactopyranosyl)-α-D-galactopyranoside (Galβ1,3GalNAc-α-pNP) in 30 min at 37°C in 100 μL 1X Reaction Buffer 7 (50 mM sodium citrate, pH 5.0). The reaction is stopped by the addition of 100 μL 0.2 M borate buffer, pH 9.8, followed by measuring absorption at 405 nm.
Supplementary Information
Form and Storage:
The product (Cat #GE1301) is supplied as a solution in 0.1 mL storage buffer (20 mM Tris-HCl, 100 mM NaCl, pH 7.5) with 10X Reaction Buffer 7 to ensure optimal digestion and ease of use. It is stable at 4°C for up to 6 months.
