PNGase A
PNGase A
- Product Code: LZ-rPNGaseA-01-S
- Size 150 units
- £311
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
Protocol | Product Specifications
SDS: PNGase A | GlycoBuffer 3| Glycoprotein Denaturing Buffer | NP-40
Product specification:
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.
Product Source: Cloned from Oryza sativa (rice) and expressed in Pichia pastoris.
Components:
| 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% |
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.
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