What is the Difference Between Nitrocellulose and PVDF?

Nitrocellulose and PVDF (polyvinylidene difluoride) are both commonly used membranes in Western blotting, but they have some differences that can impact their performance and application. Here are the main differences between the two:

1. Protein binding capacity: PVDF membranes have a higher protein binding capacity (170-200 µg/cm²) compared to nitrocellulose membranes (80-100 µg/cm²).
2. Sensitivity: PVDF membranes offer higher sensitivity, which can be beneficial for detecting low-abundance proteins but may not be ideal for high-abundance proteins due to increased background noise. Nitrocellulose membranes have lower background noise, making them suitable for proteins expressed at high levels.
3. Durability and chemical resistance: PVDF membranes are more durable and chemically resistant, making them ideal for multiple rounds of reprocessing and stripping without losing signal. Nitrocellulose membranes, on the other hand, are brittle and fragile, which can make them difficult to strip and re-probe.
4. Pore size: Both membranes come in typical pore sizes of 0.1, 0.2, and 0.45 µm, with a 0.45 µm membrane being suitable for most protein blotting applications.
5. Detection methods: Both membranes are compatible with chemiluminescence-based Western blotting. For fluorescence-based Western blotting, PVDF membranes are preferred, but low-fluorescence PVDF membranes should be used to avoid high background.
6. Protein size: PVDF membranes are better for high molecular weight proteins, while nitrocellulose membranes are better for mid to low molecular weight proteins.
7. Autofluorescence: Nitrocellulose membranes have low autofluorescence, while standard PVDF membranes have high autofluorescence, which can lead to high background.

In summary, the choice between nitrocellulose and PVDF membranes depends on your detection method, whether you intend to strip and reprobe, and your target protein(s). Nitrocellulose membranes are ideal for detecting low molecular weight proteins and high-abundance proteins, while PVDF membranes are suitable for high molecular weight proteins and low-abundance proteins.

Comparative Table: Nitrocellulose vs PVDF

Nitrocellulose and PVDF are both 'tortuous path' membranes used in Western blot analysis, but they have some differences that affect their interaction with proteins and application in Western Blot workflows. Here is a table summarizing the differences between nitrocellulose and PVDF membranes:

Both membranes can be used for Western blot analysis, but the choice between them depends on factors such as the detection method, whether you intend to strip and reprobe, and your target protein(s). Nitrocellulose membranes are ideal for detecting low molecular weight proteins, while PVDF membranes are better for detecting high molecular weight proteins.