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PNGase A

Catalog # Concentration Size List Price Quantity Your Price
P0707L 5000 units/ml 750 units $1,769.00
$1,592.10
P0707S 5000 units/ml 150 units $444.00
$399.60
Catalog # Size List Price Your Price
P0707L 750 units $1,769.00
$1,592.10
P0707S 150 units $444.00
$399.60
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

PNGase A cleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and short complex oligosaccharides such as those found in plant and insect cells from N-linked glycoproteins and glycopeptides. PNGase A differs from PNGase F in that it cleaves N-linked glycans with or without α(1,3)-linked core fucose residues.

  • Cleaves N-glycans from plant and insect derived glycoproteins and glycopeptides
  • Activity is not inhibited by an α1-3 Fucose residue on the chitobiose core
  • 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 

PNGase A is a recombinant amidase, which cleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and short complex oligosaccharides such as those found in plant and insect cells from N-linked glycoproteins and glycopeptides. PNGase A differs from PNGase F in that it cleaves N-linked glycans with or without α(1,3)-linked core fucose residues.

Substrate Specificity
PNGase A hydrolyzes N-glycan chains from glycoproteins/peptides regardless of the presence of xylose or fucose. [x = H or Man or GlcNAc].
Product Source
Cloned from Oryza sativa (rice) and expressed in Pichia pastoris.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  PNGase A P0707SVIAL 4 1 x 0.03 ml 5,000 units/ml
  GlycoBuffer 3 B1720SVIAL -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 A P0707LVIAL 4 1 x 0.15 ml 5,000 units/ml
  GlycoBuffer 3 B1720SVIAL -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 %

Properties & Usage

Unit Definition

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

Reaction Conditions

1X GlycoBuffer 3
Incubate at 37°C

1X GlycoBuffer 3
50 mM sodium acetate
(pH 6 @ 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: 63.8 kDa

Unit Assay Conditions

1 μg of recombinant Avidin is denatured with 1X Glycoprotein Denaturing Buffer at 100°C for 10 minutes. After the addition of NP-40 and GlycoBuffer 3, two-fold dilutions of PNGase A are added and the reaction mix is incubated for 1 hour at 37°C. Separation of reaction products are visualized by SDS-PAGE.

Companion Products
Notes
  • PNGase A is active on both glycoproteins and glycopeptides.
  • PNGase A cannot cleave larger N-glycans such as those from Fetuin, Fibrinogen, IgG, Lactoferrin and Transferrin.
  • PNGase A is able to cleave high mannose N-glycan structures from Man 3 up to Man 9.
  • Reactions may be scaled-up linearly to accommodate larger reaction volumes.
Citations
Additional Citations
  • Yan, S., Vanbeselaere, J., Wöls, F., et. al. (2018) Core richness of N-glycans of Caenorhabditis elegans: a case study on chemical and enzymatic release Anal Chem 90(1), 928-935.PubMedID: 29182268, DOI: DOI: 10.1021/acs.analchem.7b03898
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

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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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