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Endo Hf

Catalog # Concentration Size List Price Quantity Your Price
P0703L 1000000 units/ml 500000 units $496.00
$446.40
P0703S 1000000 units/ml 100000 units $123.00
$110.70
Catalog # Size List Price Your Price
P0703L 500000 units $496.00
$446.40
P0703S 100000 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

Endo Hf is a recombinant protein fusion of Endoglycosidase H and maltose binding protein. Endo Hf cleaves within the chitobiose core of high mannose and some hybrid oligosaccharides from N-linked glycoproteins equally as well as Endo H.

  • MBP-fusion protein for easy removal
  • 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


 

Endo Hf is a recombinant protein fusion of Endoglycosidase H and maltose binding protein. Endo Hf cleaves within the chitobiose core of high mannose and some hybrid oligosaccharides from N-linked glycoproteins (1) equally as well as Endo H.


60 μg of RNase B was incubated with 3,000 units of Endo H or Endo Hf under standard assay conditions
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-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 Hf P0703SVIAL -20 1 x 0.1 ml 1,000,000 units/ml
  GlycoBuffer 3 B1720SVIAL -20 1 x 1 ml 10 X
  Glycoprotein Denaturing Buffer B1704SVIAL -20 1 x 1 ml 10 X
  Endo Hf P0703LVIAL -20 1 x 0.5 ml 1,000,000 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 (0.5% SDS, 40 mM DTT) at 100°C for 10 minutes. After the addition of 1X GlycoBuffer 3, two-fold dilutions of Endo Hf are added and the reaction mix is incubated for 1 hour at 37°C. Separation of reaction products are 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: 70 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 the Protocol Tab.
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. Unpublished observation
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
Legal And Disclaimer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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