New England Biolabs Canada
Expand menu
Sign In or Sign UpMy NEB

PNGase F, Recombinant

Catalog # Concentration Size List Price Quantity Your Price
P0708L 500000 units/ml 75000 units $1,072.00
$964.80
P0708S 500000 units/ml 15000 units $267.00
$240.30
Catalog # Size List Price Your Price
P0708L 75000 units $1,072.00
$964.80
P0708S 15000 units $267.00
$240.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

PNGase F is the most effective enzymatic method for removing almost all N-linked oligosaccharides from glycoproteins. PNGase F, Recombinant is a recombinant amidase, which cleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides.

  • Recombinant enzyme; guaranteed endoglycosidase F1-, F2- or F3-free
  • Leaves N-glycan core oligosaccharides intact and suitable for further analysis
  • ≥ 95% purity, as determined by SDS-PAGE and intact ESI-MS
  • Stored in 50% glycerol
  • Optimal activity and stability for up to 24 months
  • Can be used under native or denaturing conditions
Featured Videos
View Video Library
  • Monarch_MiniprepProtocol_VideoThumb
    Overview of Glycobiology

Peptide: N-Glycosidase F, also known as PNGase F, is a recombinant amidase which cleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides from N-linked glycoproteins (1).
Product Source
Cloned from Elizabethkingia miricola (formerly Flavobacterium meningosepticum) and expressed in E. coli (2).
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  PNGase F, Recombinant P0708SVIAL -20 1 x 0.03 ml 500,000 units/ml
  GlycoBuffer 2 B3704SVIAL -20 1 x 1 ml 10 X
  Glycoprotein Denaturing Buffer B1704SVIAL -20 1 x 1 ml 10 X
  NP-40 B2704SVIAL -20 1 x 1 ml 10 %
  PNGase F, Recombinant P0708LVIAL -20 1 x 0.15 ml 500,000 units/ml
  GlycoBuffer 2 B3704SVIAL -20 1 x 1 ml 10 X
  Glycoprotein Denaturing Buffer B1704SVIAL -20 1 x 1 ml 10 X
  NP-40 B2704SVIAL -20 1 x 1 ml 10 %
Application Features
  • Removal of carbohydrate residues from proteins

Properties & Usage

Unit Definition

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

Unit Definition Assay:
10 µg of RNase B are denatured with 1X Glycoprotein Denaturing Buffer (0.5% SDS, 40 mM DTT) at 100°C for 10 minutes. After the addition of NP-40 and GlycoBuffer 2, two-fold dilutions of PNGase F are added and the reaction mix is incubated for 1 hour at 37°C. Separation of reaction products are visualized by SDS-PAGE.

1X Glycoprotein Denaturing Buffer
0.5% SDS
40 mM DTT

1X NP-40
1% NP-40 in MilliQ-H2O

Reaction Conditions

1X GlycoBuffer 2
Incubate at 37°C

1X GlycoBuffer 2
50 mM Sodium Phosphate
(pH 7.5 @ 25°C)

Storage Buffer

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

Heat Inactivation

75°C for 10 minutes

Molecular Weight

Apparent: 36000 daltons


Companion Products
Notes
  • To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required.
  • Since PNGase F, Recombinant activity is inhibited by SDS, it is essential to have NP-40 in the reaction mixture. It is not known why this non-ionic detergent counteracts the SDS inhibition at the present time.
  • PNGase F, Recombinant will not cleave N-linked glycans containing core α1-3 Fucose.
References
  • Maley, F. et al. (1989). Anal. Biochem. 180, 195-204.
  • Chen, M. New England Biolabs, Inc., Unpublished observation
FAQs
Tech Tips
    1. You can use this enzyme under native or denaturing conditions
    2. Under native conditions we recommend adding more enzyme and using longer incubation times
    3. PNGase F activity is inhibited by SDS, therefore under denaturing conditions it is essential to have NP-40 present in the reaction mixture in a 1:1 ratio.
    4. PNGase F will not cleave N-linked glycans containing core α1-3 Fucose (PNGase A must be used in this instance)
    5. A good positive control substrate is RNase B
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.

Other Products You May Be Interested In
 Top