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
Product Information
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 SpecificityPNGase 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.
This product is related to the following categories:
The following reagents are supplied with this product:
NEB #
Component Name
Component #
Stored at (°C)
Amount
Concentration
P0707S
4
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 %
P0707L
4
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.
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.
NEB's version of PNGase A can cleave glycoproteins, there is no need for tryptic digest prior to deglycosylation.
PNGase A can cleave N-linked glycans containing core α1-3 Fucose.
PNGase A 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.
A good positive control substrate is recombinant avidin from maize or HRP.
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
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Specifications & Change Notifications
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