PNGase F (N-Glycosidase F) is an asparagine amidase produced by Elizabethkingia miricola. PNGase F releases intact N-glycans by hydrolyzing the amide bonds of asparagine side chains (Asn-X-Ser/Thr, where X ≠ Pro), while converting Asn to aspartic acid (Asp). Unlike other glycosidases (such as Endo H), PNGase F can cleave all types of N-glycans (high-mannose, hybrid, and complex oligosaccharides), but it does not act on O-linked glycans or plant/insect N-glycans modified by core α-1,3-fucosylation. This enzyme is of significant importance in glycoprotein research, quality control in biopharmaceuticals, and proteomics. To enhance the convenience of experiments, we have developed the Fast PNGase F Kit, which simplifies the experimental procedures and reduces the reaction time to less than 20 minutes, thereby significantly improving experimental efficiency.
1. Two -Step deglycosylation
1). Add the following components to one tube on ice:
Component | Volume | Final Concentration |
Glycoprotein
| - | 10 μg/μl |
5 × Reaction Buffer | 2 μl | 1 × |
ddH2O | Up to 10 μl | - |
2). Denaturation: heating to 80℃ for 2 min
3). Deglycosylation: immediately after the reaction was completed, it was cooled on ice, and 1 μl of PNGase F was added and incubated at 50ºC for 10 minutes.
2. One -Step deglycosylation
1). Add the following components to one tube on ice:
Component | Volume | Final Concentration |
Glycoprotein | - | 10 μg/μl |
5* Reaction Buffer | 2 μl | 1 × |
PNGase F | 1 μl | - |
ddH2O | Up to 10 μl | - |
2). Deglycosylation: 1 μl of PNGase F was added and incubated at 50ºC for 10 minutes.
Two-step enzyme digestion is recommended to ensure higher enzyme digestion efficiency.
This product is for scientific research purposes only.
For your safety and health, please wear a lab coat and disposable gloves.
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