Endo F2

Catalog # Concentration Size List Price Quantity Your Price
P0772S 8000 units/ml 480 units $350.00
$315.00
Catalog # Size List Price Your Price
P0772S 480 units $350.00
$315.00
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Endo F2 is a highly specific recombinant endoglycosidase which cleaves within the chitobiose core of asparagine-linked complex biantennary and high mannose oligosaccharides from glycoproteins and glycopeptides. Endo F2 cleaves biantennary glycans at a rate approximately 20 times greater than high mannose glycans.  The activity of Endo F2 is identical on biantennary structures with and without core fucosylation.  However, Endo F2 is not active on hybrid or tri- and tetra-antennary oligosaccharides.

  • Endo F2 is tagged with a chitin binding domain (CBD) for easy removal from a reaction
  • Recombinant enzyme with no detectable exooglycosidase 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

 

Specificity:
Endo F2 Specificity
Product Source
Cloned from Elizabethkingia miricola (formerly Flavobacterium meningosepticum) and expressed in E. coli.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Endo F2 P0772SVIAL -20 1 x 0.06 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

1X 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: 39.8 kDa

Unit Assay Conditions

Two fold dilutions of Endo F2 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.

Notes
  • The enzyme can be used under denaturing and non-denaturing (native) conditions. However, optimal activity occurs under non-denaturing conditions.
  • Reactions may be scaled-up linearly to accommodate larger reaction volumes.
Tech Tips
    • Endo F2 is tagged with a chitin binding domain (CBD) for easy removal from a reaction using chitin magnetic beads (NEB #E8036 )
    • Endo F2 is typically used under non-denaturing (native) conditions, as denaturation is not necessary for optimal activity of the enzyme.
    • Concentrations of 0.5% SDS do not inhibit Endo F2 activity when used in the presence of 1% NP-40.
    • The optimal pH for cleavage of biantennary glycans is pH 4.5 (10X Glycobuffer 4)
    • Although not as optimal, cleavage can also occur at pH 6.0 (10X Glycobuffer 3).
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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