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PNGase F (Glycerol-free)

Catalog # Concentration Size List Price Quantity Your Price
P0705L 500000 units/ml 75000 units $1,299.00
$1,169.10
P0705S 500000 units/ml 15000 units $326.00
$293.40
Catalog # Size List Price Your Price
P0705L 75000 units $1,299.00
$1,169.10
P0705S 15000 units $326.00
$293.40
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 (Glycerol-free) is an amidase, which cleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides.

  • Glycerol-free for optimal performance in HPLC and mass spectrometry analysis
  • ≥ 95% purity, as determined by SDS-PAGE and intact ESI-MS
  • Non-recombinant with no detectable endoglycosidase F1, F2 or F3 contamination
  • Optimal activity and stability for up to 24 months
  • Can be used under native or denaturing conditions
  • Optimized for deglycosylation of glycoproteins; leaves N-glycan core oligosaccharides intact and suitable for further analysis
Featured Videos
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  • Monarch_MiniprepProtocol_VideoThumb
    Overview of Glycobiology

Peptide: N-Glycosidase F, also known as PNGase F, is an amidase which is supplied glycerol free for optimal performance in HPLC intensive methods. PNGase F cleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid and complex oligosaccharides from N-linked glycoproteins (1).

Detailed Specificity
PNGase F is not able to cleave N-linked glycans from glycoproteins when the innermost GlcNAc residue is linked to an α1-3 Fucose residue (2). This modification is most commonly found in plant and some insect glycoproteins.
PNGase F is not able to cleave N-linked glycans from glycoproteins when the innermost GlcNAc residue is linked to an α1-3 Fucose residue (2). This modification is most commonly found in plant and some insect glycoproteins.
Product Source
PNGase F is purified from Flavobacterium meningosepticum (3), free of contaminants (Endo F, proteases, etc.).
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  PNGase F (Glycerol-free) P0705SVIAL 4 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 (Glycerol-free) P0705LVIAL 4 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 N-linked glycans from glycoproteins
  • Preferred formulation for HPLC and MS intensive methods of glycoprotein analysis

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

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

Heat Inactivation

75°C for 10 minutes

Molecular Weight

Apparent: 36000 daltons

Storage Notes

  • Do not freeze


Companion Products
Notes
  • Since PNGase F activity is inhibited by SDS, it is essential to have NP-40 present in the reaction mixture. Why this non-ionic detergent counteracts the SDS inhibition is unknown at present.
  • To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required. 
  • PNGase F will not cleave N-linked glycans containing core α1-3 Fucose. 
  • Repeated freeze thaw cycles degrade enzyme activity over time. 
  • Typical reaction conditions: Please see Protocol tab
References
  • Maley, F. et al. (1989). Anal. Biochem.. 180, 195-204.
  • Tretter, V. et al. (1991). Eur. J. Biochem.. 199, 647-652.
  • Plummer, T.H. Jr. and Tarentino, A.L. (1991). Glycobiology. 1, 257-263.
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
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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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