Endo H cleaves asparagine-linked hybrid or high mannose oligosaccharides but not complex oligosaccharides
Product Description Endo H, endo-beta-N-acetylglucosaminidase H, Endoglycosidase H Endo H cleaves Asparagine-linked hybrid or high mannose oligosaccharides, but not complex oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Detergent and heat denaturation may increase the rate of cleavage for some glycoproteins. Source recombinant from Streptomyces plicatus in E.Coli
EC 3.2.1.96 Specific Activity >40 U/mg Activity >5 U/ml Molecular weight 29,000 daltons pH range 5-6, optimum 5.5
Specific Activity One unit of Endo H activity is defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 µmole of denatured Ribonuclease B. Cleavage is monitored by SDS-PAGE (cleaved Ribonuclease B migrates faster).
Contents 60 µl aliquot of enzyme (300 mU) in 20 mM Tris-HCl, 25 mM NaCl, 1 mM EDTA (pH 7.5). Included with 20 µL and 60 µL pack sizes: 200 µl 5x Reaction Buffer 5.5 (250 mM sodium phosphate, pH 5.5) Denaturation Solution – 2% SDS, 1 M Beta-mercaptoethanol
Suggested usage 1. Add up to 200 µg of glycoprotein to Eppendorf tube. Adjust to 37.5 µl final volume with deionized water. 2. Add 10 µl 5X Endo H Buffer and 2.5 µl of Denaturation Solution (SDS/-ME). Heat at 100˚C for 5 minutes. 3. Cool, and then add 2.0 µl of Endo H to the reaction. Incubate 3 hours at 37˚C.
Storage Store enzyme at 4˚C. Stability Stable at least 12 months when stored properly. Several days exposure to ambient tempertures will not reduce activity. Purity Endoglycosidase H is tested for contaminating protease as follows; 10 μg of denatured BSA is incubated for 24 hours at 37oC with 2 μL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation.
The production host strain has been extensively tested and does not produce any detectable glycosidases. | 
