Product Class: Other

Bacteroides Heparinase II
NEBU cloned at NEB recombinant 30 100 Heat

Product Introduction

Bacteroides Heparinase II (also called Heparin Lyase II) is cloned from Bacteroides eggerthii. It is a low specificity enzyme that is active on both heparin and heparan sulfate.

Bacteroides Heparinase II cleaves the glycosidic bond between N-sulfated and glucuronic or iduronic acid residues. The reaction yields oligosaccharide products containing unsaturated uronic acids which can be detected by UV spectroscopy at 232 nm.

When used alone this enzyme rarely yields complete depolymerization of a polysaccharide chain, however disaccharide analysis is enhanced when used in combination with Heparinase I and III.

• Recombinant enzyme with no detectable glycosidase, sulfatase or uronidase contaminating activities
• ≥95% purity, as determined by SDS-PAGE and intact ESI-MS
• Optimal activity and stability for up to 12 months when stored in solution at -80°C

 

Catalog # Size Concentration
P0736S 80.0 units 4000 units/ml
P0736L 200.0 units 4000 units/ml

Product Information

Description




Product Source

Cloned from Bacteroides Eggerthii and expressed in E. coli.
This product is related to the following categories:
Heparinases Products
This product can be used in the following applications:
Analysis of Heparin/HS

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • P0736S     -80    
      Bacteroides Heparinase II P0736SVIAL -80 1 x 0.02 ml 4,000 units/ml
      Bacteroides Heparinase Reaction Buffer B0735SVIAL -20 1 x 1 ml 10 X
  • P0736L     -80    
      Bacteroides Heparinase II P0736LVIAL -80 1 x 0.05 ml 4,000 units/ml
      Bacteroides Heparinase Reaction Buffer B0735SVIAL -20 1 x 1 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will liberate 1.0 μmol unsaturated oligosaccharides from porcine mucosal heparin per minute at 30°C and pH 7.0 in a total reaction volume of 100 μl.

Reaction Conditions

1X Bacteroides Heparinase Reaction Buffer
Incubate at 30°C

Storage Buffer

20 mM Tris-HCl
100 mM NaCl
1 mM EDTA
5 mM CaCl2
pH 7.5 @ 25°C

Heat Inactivation

100°C for 1 minute

Molecular Weight

Apparent: 86 kDa

Unit Assay Conditions

Two fold dilutions of Bacteroides Heparinase II are incubated with 1 mg/ml porcine mucosal heparin substrate in 1X Bacteroides Heparinase Reaction Buffer, in a 100 μl reaction. The reaction mix is incubated at 30°C. Liberation of unsaturated oligosaccharides is detected by real-time UV spectroscopy at 232 nm.

Product Notes

  1. Avoid repeated freeze-thaw cycles.
  2. Reaction Conditions:
    1. Combine 10 μl of 1 mg/ml heparin substrate, 10 μl Bacteroides Heparinase Reaction Buffer and H2O in a total reaction volume of 100 μl. 
    2. Add 1 μl Bacteroides Heparinase II.
    3.  Incubate reaction at 30°C for 1–24 hours (monitor absorbance at 232 nm for determination of partial or complete digestion). 
    Optimal incubation times and enzyme concentrations must be determined empirically for a particular substrate.

FAQs & Troubleshooting

FAQs

  1. What are Bacteroides Heparinase Enzymes?
  2. What is the difference between Bacteroides Heparinase I, II and III?
  3. Can Bacteroides Heparinase I, II, and III be used together in one digest?
  4. What is the optimal pH range for Bacteroides Heparinase II?
  5. Is Bacteroides Heparinase II activated by Calcium?

Tech Tips

Heparinase II has a broad specificity and cleaves domains of both high and low sulfation on both heparin and HS.

Bacteroides Heparinase II is most active between pH 7.0 - 8.0.

Bacteroides Heparinase II is calcium dependent and has greatest activity in the presence of 1.5 - 5mM CaCl2.

Bacteroides Heparinase II can be used in double digests with Bacteroides Heparinase I and III using the Heparinase reaction buffer.

Quality, Safety & Legal

Quality Assurance Statement

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 DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

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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