Product Overview
PNGase F is the most efficient enzymatic method to remove nearly all N-glycans from glycoproteins. PNGase F digests the glycosidic bond between the innermost N-acetylglucosamine and asparagine residues. It cleaves N-glycans attached to glycoproteins, including high-mannose, hybrid, and complex oligosaccharide glycoforms.
Key Features
Efficient N-glycan release enzyme
PNGase F-Fast enables enzymatic removal of N-linked glycans from glycoproteins by cleaving the bond between the innermost N-acetylglucosamine and asparagine residues. It is applicable to high-mannose, hybrid, and complex N-glycan structures.
Preservation of glycan core structure
After enzymatic digestion, released N-glycans retain their core structures, supporting downstream analytical applications.
Rapid reaction setup
Supports fast deglycosylation workflows with a typical setup time of approximately 10 minutes at 50 °C under recommended conditions.
High-concentration formulation
High-concentration design allows flexible adjustment of enzyme input in different sample types and reaction scales, supporting more demanding or high-load glycoprotein workflows.
High purity
Purity is typically ≥95% as determined by standard analytical methods.
Flexible reaction compatibility
Compatible with both denaturing and non-denaturing digestion conditions depending on experimental requirements.
Multiple product formats available
Different formats are available to support diverse application needs (e.g., HBP003006 / HBP003004 / HBP003005).
Applications
N-glycan release for analytical workflows
Used for enzymatic removal of N-linked glycans in workflows typically performed at 50 °C for downstream structural analysis.
CE and LC/LC-MS-based glycan analysis
Suitable for sample preparation in capillary electrophoresis (CE), HPLC, and LC-MS-based N-glycan profiling applications.
Glycoprotein modification for downstream applications
Applied in workflows requiring removal of N-glycans prior to protein modification or bioconjugation processes, including ADC-related research applications depending on experimental design.
Performance Data
Digestion efficiency
Samples of IgG1, IgG4 and fusion protein were digested with PNGase F at 50 °C for 5 min, followed by CE-SDS analysis to assess digestion efficiency.
| Sample | Protein content | PNGase F Volume |
Buffer Volume |
Tem. | Digestion Time |
Reaction Volume |
Digestion Efficiency |
|---|---|---|---|---|---|---|---|
| IgG1 | 40 ug | 1.2 μL | 3 μL | 50℃ | 5min | 24.2 μL | 100% |
| IgG4 | 40 ug | 1.2 μL | 3 μL | 50℃ | 5min | 24.2 μL | 100% |
| Fusion Protein | 40 ug | 1.2 μL | 3 μL | 50℃ | 5min | 24.2 μL | 100% |
FAQ
What components are included in the PNGase F (High Concentration) system?
The system typically includes PNGase F (high-concentration formulation) and PNGase F buffer, designed to support flexible enzyme loading in N-linked glycan release workflows.
What is the advantage of the high-concentration formulation?
The high-concentration formulation allows flexible adjustment of enzyme input depending on sample type, glycoprotein content, and reaction scale, making it suitable for workflows requiring higher enzyme loading or optimized reaction efficiency.
Can this system be used under both denaturing and non-denaturing conditions?
Yes. PNGase F (High Concentration) is compatible with both denaturing and non-denaturing digestion conditions, depending on experimental design and sample characteristics.
What are the recommended reaction conditions?
The system supports rapid deglycosylation workflows, typically performed at 50 °C with a short setup time (approximately 10 minutes) under recommended conditions, followed by incubation according to the specific protocol and sample type.
Can reaction conditions be optimized?
Yes. Enzyme amount, incubation time, and reaction conditions can be adjusted based on glycoprotein complexity, sample load, and downstream analytical requirements.
Protocols under denaturing and non-denaturing conditions
| Steps | Denaturing Reaction Conditions | Non-Denaturing Reaction Conditions |
|---|---|---|
| 1 | Dissolve 40~100 µg of antibody or glycoprotein with deionized water to a final concentration of 2~5 mg/mL, then add 20 μL of the sample. | Dissolve 40~100 µg of antibody or glycoprotein with deionized water to a final concentration of 2~5 mg/mL, then add 20 μL of the sample. |
| Add 3µL of PNGase F buffer | Change the buffer to 0.25M PB (pH 7.5~8.0) | |
| 2 | 90℃ 3min | / |
| 3 | Add 1.2µL of PNGaseF | Add 1.2µL of PNGaseF |
| 5 | 50℃ 5min | 50℃≥0.5h or 37℃≥4h （can be adjusted if needed） |
