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Endo H

Catalog # Concentration Size List Price Quantity Your Price
P0702L 500000 units/ml 50000 units $496.00
$446.40
P0702S 500000 units/ml 10000 units $123.00
$110.70
Catalog # Size List Price Your Price
P0702L 50000 units $496.00
$446.40
P0702S 10000 units $123.00
$110.70
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Endoglycosidase H is a recombinant glycosidase which cleaves within the chitobiose core of high mannose and some hybrid oligosaccharides from N-linked glycoproteins.

  • 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


 
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

60 mg 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
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 picatus (2) and overexpressed in E.coli (3).
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  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
  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
Application Features
  • Removal of high mannose N-glycans from glycoproteins

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

Reaction Conditions

1X GlycoBuffer 3
Incubate at 37°C

1X GlycoBuffer 3
50 mM sodium acetate
(pH 6 @ 25°C)

Storage Buffer

20 mM Tris-HCl
50 mM NaCl
5 mM EDTA
pH 7.5 @ 25°C

Heat Inactivation

75°C for 10 minutes

Molecular Weight

Apparent: 29 kDa


Companion Products
Notes
  • Enzymatic activity is not affected by SDS.
  • 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.
Citations
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
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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