Endoglycosidase H is a recombinant glycosidase which cleaves within the chitobiose core of high mannose and some hybrid oligosaccharides from N-linked glycoproteins. A tagged version, which is a recombinant protein fusion of Endoglycosidase H and maltose binding protein is also available, Endo Hf (NEB #P0703).
Recombinant enzyme with no detectable exoglycosidase or other endoglycosidase contaminating activities
Glycerol-free for optimal performance in HPLC and mass spectrometry analysis
≥95% purity, as determined by SDS-PAGE and intact ESI-MS
Optimal activity and stability for up to 24 months
Product Information
Endoglycosidase H is a recombinant glycosidase which cleaves within the chitobiose core of high mannose and some hybrid oligosaccharides from N-linked glycoproteins.
60 µg of RNase B was incubated with 3,000 units of Endo H or Endo Hf under standard assay conditions
Aliquots were removed at various time points and measured for released carbohydrate. [Dubois et al. (1956) Anal. Chem. 28, 350-356].Mobility Shift Analysis 1 unit of Endo H, Endo Hf or PNGase F was incubated per 10 µg of RNase B under standard assay conditions. At various time points, aliquots were removed and analyzed on a 10-20% SDS-PAGE gel for carbohydrate (CHO) release. 1 unit is defined as the amount of enzyme required to remove >95% of the carbohydrate from 10 µg of RNase B in one hour at 37°C.
Product Source
Cloned from Streptomyces plicatus (2) and overexpressed in E.coli (3).
This product is related to the following categories:
The following reagents are supplied with this product:
NEB #
Component Name
Component #
Stored at (°C)
Amount
Concentration
P0702S
-20
Endo H
P0702SVIAL
-20
1 x 0.02 ml
500,000 units/ml
GlycoBuffer 3
B1720SVIAL
-20
1 x 1 ml
10 X
Glycoprotein Denaturing Buffer
B1704SVIAL
-20
1 x 1 ml
10 X
P0702L
-20
Endo H
P0702LVIAL
-20
1 x 0.1 ml
500,000 units/ml
GlycoBuffer 3
B1720SVIAL
-20
1 x 1 ml
10 X
Glycoprotein Denaturing Buffer
B1704SVIAL
-20
1 x 1 ml
10 X
Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 µg of denatured RNase B in 1 hour at 37°C in a total reaction volume of 10 µl (10 NEB units = 1 IUB milliunit).
Unit Definition Assay: 10 µg of RNase B are denatured with 1X Glycoprotein Denaturing Buffer at 100°C for 10 minutes. After the addition of 1X GlycoBuffer 3, two-fold dilutions of Endo H are added and the reaction mix is incubated for 1 hour at 37°C. Separation of reaction products is visualized by SDS-PAGE.
1X Glycoprotein Denaturing Buffer 0.5% SDS 40 mM DTT
To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required.
Activity at different temperatures (measured after a 1 hour incubation of glycosidase and denatured RNase B at the given temperature): 37°C - 100%; 30°C - 65%; 25°C - 40%; 17°C - 25% and 2°C - 0%.
Typical reaction conditions: Please see FAQs.
References
Maley, F. et al. (1989). Anal. Biochem. 180, 195-204.
Robbins, P. et al. (1984). J. Biol. Chem. 259, 7577-7583.
Guan, C. and Wong,S. New England Biolabs, unpublished observations.
You can use this enzyme under native or denaturing conditions
Under native conditions we recommend adding more enzyme and using longer incubation times
Enzymatic activity is not affected by SDS
A good positive control substrate is RNase B
Citations & Technical Literature
Citations
Product Citation Tool
Additional Citations
Arakel EC, Brandenburg S, Uchida K, Zhang H, Lin YW, Kohl T, Schrul B, Sulkin MS, Efimov IR, Nichols CG, Lehnart SE, Schwappach B (2014) Tuning the electrical properties of the heart by differential trafficking of KATP ion channel complexes J Cell Sci; 127(Pt 9), 2106-19. PubMedID: 24569881, DOI: 10.1242/jcs.141440
Möykkynen T, Coleman SK, Semenov A, Keinänen K (2014) The N-terminal domain modulates α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor desensitization J Biol Chem; 289(19), 13197-205. PubMedID: 24652293, DOI: 10.1074/jbc.M113.526301
Rosenbaum EE, Vasiljevic E, Brehm KS, Colley NJ (2014) Mutations in four glycosyl hydrolases reveal a highly coordinated pathway for rhodopsin biosynthesis and N-glycan trimming in Drosophila melanogaster PLoS Genet; 10(5), e1004349. PubMedID: 24785692, DOI: 10.1371/journal.pgen.1004349
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Specifications & Change Notifications
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