Endoglycosidase H

- Overview
  - Endo H cleaves asparagine-linked hybrid or high mannose oligosaccharides but not complex oligosaccharides.
    
    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.
    
    Contents:
    60 µL aliquot of enzyme (300 mU) in 20 mM Tris-HCl, 25 mM NaCl, 1 mM EDTA (pH 7.5).
    200 µL 5x Reaction Buffer 5.5 (250 mM sodium phosphate, pH 5.5)
    Denaturation Solution - 2% SDS, 1 M Beta-mercaptoethanol
    
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    More Details
    
    Glycosidases
- Properties
  - Name
    
    Endo H, endo-β-N-acetylglucosaminidase H, Endoglycosidase H
    
    Purification
    
    Endoglycosidase H is tested for contaminating protease as follows; 10 µg of denatured BSA is incubated for 24 hours at 37°C with 2 µL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation.
    
    Storage
    
    Store enzyme at 4°C.
    
    Note
    
    Stable at least 12 months when stored properly. Several days exposure to ambient tempertures will not reduce activity.
    
    Activity
    
    >5 U/mL
    
    More Information
    
    Database Link
    
    EC 3.2.1.96
    
    * For research use only
- Applications
  - Application
    
    Glycan Release
- Reference
  - 1. Robbins P. W., D. F. Wirth and C. J. Hering. Expression of the Streptomyces enzyme endoglycosidase H in Escherichia coli. Biol Chem 256:10640-10644 (1981).
    2. Robbins P. W., R. B. Trimble, D. F. Wirth, C. Hering, F. Maley , G. F. Maley, R. Das, B. W. Gibson, N. Royal and K. Biemann. Primary structure of the Streptomyces enzyme endo-beta-N-acetylglucosaminidase H. J Biol Chem 259:7577-7583 (1984).
    3. Trimble R. B., A. L. Tarentino , G. E Aumick and F. Maley. Endo-beta-N-acetylglucosaminidase L from Streptomyces plicatus. Methods Enzymol 83:603-610 (1982).
    4. Trimble R. B. and F. Maley. Optimizing hydrolysis of N-linked high-mannose oligosaccharides by endo-beta-N-acetylglucosaminidase H. Anal Biochem 141:515-522 (1984).
    5. Trimble R. B., R. J. Trumbly and F. Maley. Endo-beta-N-acetylglucosaminidase H from Streptomyces plicatus. Methods Enzymol 138:763-770 (1987).
    6. Trumbly R. J., P. W. Robbins, M. Belfort, F. D. Ziegler, F. Maley and R. B. Trimble. Amplified expression of Streptomyces endo-beta-N-acetylglucosaminidase H in Escherichia coli and characterization of the enzyme product. J Biol Chem 260:5683-5690 (1985).

Documents

Specsheet

CofA

MSDS

Endoglycosidase H

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