Product Class: Other

Endo F3
NEBU cloned at NEB recombinant 37 65 Heat

Product Introduction

Endo F3 is an endoglycosidase that cleaves within the chitobiose core of N-linked fucosylated-biantennary and triantennary complex oligosaccharides from glycoproteins. Endo F3 is tagged with chitin binding domain (CBD) 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
Catalog # Size Concentration
P0771S 240.0 units 8000 units/ml

Product Information

Description

Endo F3 is a highly specific recombinant endoglycosidase which cleaves within the chitobiose core of asparagine-linked fucosylated-biantennary and triantennary complex oligosaccharides from glycoproteins. Endo F3 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.

Specificity
P0771 specificity

Product Source

Cloned from Elizabethkingia miricola (formerly Flavobacterium meningosepticum) and expressed in E. coli.
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
  • P0771S     -20    
      Endo F3 P0771SVIAL -20 1 x 0.03 ml 8,000 units/ml
      GlycoBuffer 4 B1703SVIAL -20 1 x 1 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave > 95% of the carbohydrate from 10 µg Porcine Fibrinogen in 1 hour at 37°C in a total reaction volume of 10 µl.

Reaction Conditions

1X GlycoBuffer 4
Incubate at 37°C

GlycoBuffer 4
50 mM sodium acetate
(pH 4.5 @ 25°C)

Storage Buffer

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

Heat Inactivation

65°C for 10 minutes

Molecular Weight

Apparent: 38.8 kDa

Unit Assay Conditions

Two fold dilutions of Endo F3 are incubated with 10 µg Porcine Fibrinogen and 1X GlycoBuffer 4 in a 10 µl reaction. The reaction mix is incubated at 37°C for 1 hour. Separation of reaction products are visualized via SDS-PAGE.

Product Notes

  1. The optimal pH for the cleavage of fucosylated-biantennary (i.e. porcine fibrinogen) and triantennary glycans (i.e. fetuin) is pH 4.5 (10X GlycoBuffer 4). However, fucosylated-biantennary glycans can also be cleaved at pH 6.0 (10X GlycoBuffer 3).
  2. The enzyme should be used under non-denaturing conditions. Detergents, such as SDS, inhibit Endo F3 activity.
  3. Reactions may be scaled-up linearly to accommodate larger reaction volumes.

Protocols, Manuals & Usage

Protocols

  1. Typical Reaction Conditions for Endo F3 Protocol (P0771)

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Is Endo F3 tagged?  
  2. What is the difference between PNGase F and Endo F3?
  3. How much Endo F3 should I use to deglycosylate a glycoprotein under native conditions?
  4. What is the preferred substrate for Endo F3?
  5. Do detergents inhibit Endo F3 activity?
  6. What is the optimal pH for Endo F3 activity?
  7. How do I eliminate Endo F3 from a reaction?
  8. What is the binding capacity of the Magnetic Chitin Beads used to remove Endo F3?
  9. Is Endo F3 compatible with downstream analysis such as HPLC and Mass Spectrometry?
  10. What are Glycosidases and their uses?

Tech Tips

  • Endo F3 is tagged with a chitin binding domain (CBD) for easy removal from a reaction using chitin magnetic beads (NEB #E8036)
  • Endo F3 should be used under non-denaturing conditions. Detergents, such as SDS, inhibit Endo F3 activity.
  • The optimal pH for the cleavage of triantennary glycans (i.e. fetuin) is pH 4.5 (10X Glycobuffer 4).
  • Fucosylated-biantennary glycans (i.e. porcine fibrinogen) can be cleaved at both pH 4.5 (10X Glycobuffer 4) and at pH 6.0 (10X Glycobuffer 3).

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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