Product Class: Other

Endo S
NEBU cloned at NEB recombinant 37 55 Heat

Product Introduction

Endo S is an endoglycosidase specific for cleaving the N-linked glycans from the chitobiose core of the heavy chain of native IgG

  • Active on native IgG
  • Tagged with a chitin binding domain (CBD) for easy removal
  • Recombinant enzyme with no detectable exooglycosidase or other endoglycosidases 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 12 months

Catalog # Size Concentration
P0741S 6000.0 units 200000 units/ml
P0741L 30000.0 units 200000 units/ml

Product Information

Description


 
Endo S is an endoglycosidase with a uniquely high specificity for removing N-linked glycans from the chitobiose core of the heavy chain of native IgG (1). Endo S is tagged with a chitin binding domain (CBD) for easy removal from a reaction and is supplied glycerol free for optimal performance in HPLC and MS intensive methods.

Product Source

Endo S is cloned from Streptococcus pyogenes and overexpressed as a fusion to the chitin binding domain in E. coli.

Specificity

Detailed Specificity
X = protein, peptide, Asparagine, or free glycan, as Endo S does not have a strict peptide requirement for activity. Endo S is active on a substrate with or without core α(1-6)fucosylation as well as a with or without a bisecting N-acetylglucosamine. Triantennary and tetrantennary sialyated or asialo glycans are not a substrate for Endo S (1).
This product is related to the following categories:
Endoglycosidases Products,
Proteome Analysis Products,
This product can be used in the following applications:
Expression Systems,
Glycan Sequencing,
Proteomics,
Recombinant Glycoprotein Expression,
Glycoprotein Analysis

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • P0741S     4    
      Endo S P0741SVIAL 4 1 x 0.03 ml 200,000 units/ml
      GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X
  • P0741L     4    
      Endo S P0741LVIAL 4 1 x 0.15 ml 200,000 units/ml
      GlycoBuffer 1 B1727SVIAL -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 5 μg of native mouse monoclonal IgG in 1 hour at 37°C in a total reaction volume of 10 µl.

Reaction Conditions

1X GlycoBuffer 1
Incubate at 37°C

1X GlycoBuffer 1
5 mM CaCl2
50 mM sodium acetate
(pH 5.5 @ 25°C)

Storage Buffer

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

Heat Inactivation

55°C for 10 minutes

Molecular Weight

Apparent: 136000 daltons

Unit Assay Conditions

5 µg of IgG in 1X GlycoBuffer 1 are incubated with two-fold dilutions of Endo S for 1 hour at 37°C. Separation of reaction products is visualized by SDS-PAGE.

Product Notes

  1. Recommended storage temperature is 4°C,avoid repeat freeze-thaw cycles.
  2. Removal of Endo S from thedeglycosylation reaction can be scaled uplinearly with larger volumes of chitin magneticbeads.
  3. The binding capacity of Chitin Magnetic Beads is 1 mg of CBD-tagged protein / mL of beads.

References

  1. Bielik, A., McLeod, E. and Magnelli, P., New England Biolabs,Inc., unpublished results.. Unpublished observation

Protocols, Manuals & Usage

Protocols

  1. Reaction Conditions for Endo S (P0741)
  2. Endo S Removal Magnetic Chitin Bead Protocol (P0741)

Application Notes

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Is Endo S tagged?
  2. What is the difference between PNGase F and Endo S?
  3. How much Endo S should I use to deglycosylate a glycoprotein under native conditions?
  4. What is the preferred substrate for Endo S?
  5. What are the typical reaction conditions for Endo S?
  6. How do I eliminate Endo S from a reaction?
  7. What is the binding capacity of the Magnetic Chitin Beads used to remove Endo S?
  8. Is Endo S compatible with downstream analysis such as HPLC and Mass Spectrometry?
  9. Why have the NEB Glycosidase enzymes changed reaction buffers? What are the new reaction buffers and can I still use an enzyme with its old buffer? Where can I find the composition of the old buffers?
  10. What are Glycosidases and their uses?

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.

Trademarks

NEW ENGLAND BIOLABS® is a registered trademark of New England Biolabs, Inc.
REMOVE-IT™ is a trademark of New England Biolabs, Inc.

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