Endo H (Endoglycosidase H) is a recombinant glycosidase expressed and purified from E.coli which carries the Streptomyces plicatus gene.
Endo H cleaves between the N-acetylglucosamine residues of the chitobiose core of N-linked glycans, leaving one N-acetyl glucosamine residue attached to the asparagine.Endo H has a molecular weight of ~31.1 kDa.The workable pH range is between 5.0 to 6.0, with the optimal pH at 5.5.Substrate Specificity- High mannose oligosaccharides- Hybrid N-linked oligosaccharidesProduct Component- Endo H : 100 ㎕ (100 unit/㎕)- 10x Reaction Buffer : 500 mM sodium citrate, pH 5.5.Unit Definition- One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 ㎍ of denatured RNase B for 1 hr at 37°C in a total reaction volume of 10 ㎕.Specific Activity- 500 unit/㎕, 400-500 unit/㎍Storage Buffer- 20 mM Tris-HCl, pH 7.5, 50 mM NaCl, 5 mM EDTA, 50% glycerol.Endo H Reaction Condition- 10 ㎍ of RNase B is denatured with 0.5% SDS, 1% β-mercaptoethanol at 100°C for 10 min. After the addition of Reaction Buffer to final 1x, Endo H are added and the reaction mix is incubated for 1 hr at 37°C. Separation of reaction products is analyzed by SDS-PAGE (cleaved RNase B migrates faster). The activity of Endo H is not affected by addition of 0.5% SDS. For a native glycoprotein, 5-fold more enzyme and longer incubation time will be required.Storage Temperature- Store at -20℃.QC Tests- Activity, SDS-PAGE purity, performance tests.
