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Specific Activity >40 U/mg;Activity >5 U/ml for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
Store at 2-8°C Ships Wet ice Check lot-specific COA for exact specifications.
SDS, COA, datasheet, and spec sheet available for download. Lot-specific COA accessible via lot number lookup.
Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
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.
Comprehensive hazard, handling, storage, and regulatory compliance document.
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