Product Class: Other

Endoglycoceramidase I (EGCase I)
NEBU cloned at NEB recombinant 37 65 Heat

Product Introduction

  • Catalyzes the hydrolysis of the β-glycosidic linkage between oligosaccharides and ceramides in various glycosphingolipids
  • Oligosaccharide and ceramide remain intact, leaving them suitable for further analysis. 
  • This is an Enzyme for Innovation (EFI). EFI is a project initiated by NEB to provide unique enzymes to the scientific community in the hopes of enabling the discovery of new and innovative applications. These enzymes have interesting properties and unique specificities.
Catalog # Size Concentration
P0773S 150.0 milliunits 6 units/ml

Product Information

Description

Endoglycoceramidase I (EGCase I) catalyzes the hydrolysis of the β-glycosidic linkage between oligosaccharides and ceramides in various glycosphingolipids. The enzyme leaves the oligosaccharide and ceramide intact, keeping them suitable for further analysis. The enzyme efficiently releases oligosaccharides from ganglio- and globo-type glycosphingolipids, certain type of cerebrosides (glucosylcerebrosides), and exhibits low activity towards galactocerebrosides (See Reference 1, FAQ #3). EGCase I does not hydrolyze sulfatides nor psychosine.

Product Source

Cloned from Rhodococcus triatomea and expressed in E. coli.
This product is related to the following categories:

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • P0773S     -20    
      Endoglycoceramidase I (EGCase I) P0773SVIAL -20 1 x 0.025 ml 6 units/ml
      EGCase I Reaction Buffer B0773SVIAL -20 1 x 1.5 ml 10 X

Properties & Usage

Unit Definition

One unit of R. triatomea EGCase I was defined as the amount of enzyme required to hydrolyze 1 μmol of ganglioside GM1a per minute at 37°C.

Reaction Conditions

1X EGCase I Reaction Buffer
Incubate at 37°C

1X EGCase I Reaction Buffer
50 mM sodium acetate
0.1% Triton® X-100
(pH 5.2 @ 25°C)

Storage Buffer

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

Heat Inactivation

65°C for 20 minutes

Molecular Weight

Apparent: 50 kDa

Unit Assay Conditions

Two fold dilutions of EGCase I are incubated with 10 nmoles of GM1a substrate in 1X EGCase I Reaction Buffer in a 10 µl reaction. The reaction mix is incubated for 5 minutes at 37°C. Released glycans are labeled with 2-AB, and reaction products are analyzed by ultra-performance hydrophilic interaction liquid chromatography with fluorescence detection.

References

  1. Albrecht, S. et al. (2016). Analytical Chemistry. 88, 4795-4802.

Protocols, Manuals & Usage

Protocols

  1. Protocol for use with EGCase I (P0773)

FAQs & Troubleshooting

FAQs

  1. Can the enzyme be stored at other temperatures?
  2. How do I optimize the reaction?
  3. What additional substrates are hydrolyzed by EGCase I?
  4. Can I purchase large amounts of an existing Enzyme for Innovation?
  5. What are Enzymes for Innovation?

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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