PVDF vs Nitrocellulose: Complete Comparison Guide
Choosing between PVDF (Polyvinylidene Difluoride) and nitrocellulose membranes is one of the most important decisions in western blotting. Both membranes bind proteins, but they differ in protein binding capacity, mechanical strength, compatibility with detection methods, and cost. This comprehensive guide provides detailed comparisons of both membrane types, including their advantages, disadvantages, protocols, and recommendations for when to use each membrane.
Overview
Protein transfer membranes are essential components of western blotting that bind proteins after transfer from the gel. Two main membrane types are used:
- PVDF: Synthetic polymer membrane with high protein binding capacity
- Nitrocellulose: Natural polymer membrane with good protein binding
Key differences:
- Protein binding: PVDF has higher binding capacity
- Mechanical strength: PVDF is more durable
- Compatibility: Both work with chemiluminescence and fluorescence
- Cost: Nitrocellulose is generally less expensive
- Handling: PVDF requires methanol activation
- Stripping: Both can be stripped, but PVDF is more resistant
The choice between membranes depends on your protein binding needs, detection method, budget, and experimental requirements.
PVDF Membrane
Characteristics
PVDF is a synthetic polymer membrane with high protein binding capacity and excellent mechanical strength. It requires activation with methanol before use.
- High protein binding capacity (150-200 μg/cm²)
- Excellent mechanical strength and durability
- Requires methanol activation before use
- Compatible with both chemiluminescence and fluorescence
- Can be stripped and reprobed multiple times
- Stable for long-term storage
Advantages
- High binding capacity: Binds more protein than nitrocellulose
- Durable: Strong and resistant to tearing
- Stripping: Can be stripped and reprobed multiple times
- Long-term storage: Stable for archival purposes
- Low background: Generally low background signal
- Good for low abundance proteins: High binding capacity helps
Disadvantages
- Requires activation: Must be activated with methanol
- Higher cost: More expensive than nitrocellulose
- Brittle when dry: Can crack if allowed to dry completely
- Hydrophobic: Requires methanol for wetting
Nitrocellulose Membrane
Characteristics
Nitrocellulose is a natural polymer membrane with good protein binding capacity. It is hydrophilic and does not require activation.
- Good protein binding capacity (80-100 μg/cm²)
- Hydrophilic, wets easily with water
- No activation required
- Compatible with both chemiluminescence and fluorescence
- Can be stripped, but less durable than PVDF
- Less expensive than PVDF
Advantages
- Easy to use: No activation required
- Cost-effective: Less expensive than PVDF
- Hydrophilic: Wets easily with water
- Good for routine use: Suitable for most applications
- Flexible: Less brittle than PVDF
Disadvantages
- Lower binding capacity: Binds less protein than PVDF
- Less durable: More fragile than PVDF
- Limited stripping: Can be stripped but less resistant
- Storage: Less stable for long-term storage
Side-by-Side Comparison
| Feature | PVDF | Nitrocellulose |
|---|---|---|
| Protein Binding Capacity | High (150-200 μg/cm²) | Good (80-100 μg/cm²) |
| Mechanical Strength | Excellent | Good |
| Activation Required | Yes (methanol) | No |
| Cost | Higher | Lower |
| Stripping/Reprobing | Excellent | Good |
| Long-term Storage | Excellent | Good |
| Ease of Use | Moderate (requires activation) | Easy |
Selection Guide
Choose PVDF When:
- Detecting low abundance proteins
- Need maximum protein binding capacity
- Plan to strip and reprobe multiple times
- Need durable membrane for handling
- Long-term storage is required
- Budget allows for higher cost
Choose Nitrocellulose When:
- Routine western blotting
- Cost is a concern
- Want simple, easy-to-use membrane
- Detecting moderate to high abundance proteins
- Single detection (no reprobing needed)
- Standard applications
General Recommendations
- For most applications: Nitrocellulose is sufficient and cost-effective
- For low abundance: PVDF offers better binding capacity
- For reprobing: PVDF is more durable
- For beginners: Nitrocellulose is easier to use
- For routine use: Choose based on budget and needs