PVDF and Nitrocellulose: Which Shou

PVDF and Nitrocellulose: Which Should You Use?

When it comes to Western blotting, most researchers prefer polyvinylidene difluoride (PVDF) and nitrocellulose over other types of membranes since both offers several key attributes that makes them suitable for particular experimental conditions. How does one compare with the other? And how do you choose which one to use in your application? To do this, you need to take the membrane type, pore size and membrane format into consideration to help you choose which membrane to use.

Protein binding capacity. PVDF has a protein binding capacity of 170 to 200 μg/cm2 while nitrocellulose has a protein binding capacity of 80 to 100 μg/cm2. Since PVDF has a higher protein binding capacity, it also offers higher sensitivity. While this feature allows it to detect lowly expressed proteins, you are more likely to get higher background noise in your antibody detection steps when using this membrane. Nitrocellulose membrane, on the other hand, may not be capable of the detection sensitivity of PVDF membranes but it will produce a lower background noise.

Binding interactions. Protein molecules bind to nitrocellulose membranes through hydrophobic interactions while molecules bind to PVDF membranes through hydrophobic and dipole interactions. However, please note that nitrocellulose requires the use of methanol in the transfer buffer which may reduce the pore size of the gel and cause high molecular weight proteins to precipitate.

Physical characteristics. While nitrocellulose is brittle and fragile, PVDF is more durable and has higher chemical resistance making it ideal for reprobing and sequencing applications. Nitrocellulose can prove to be difficult to strip and reprobe without losing signal. However, supported versions of nitrocellulose membranes are considerably more durable and resilient than standard nitrocellulose membranes so they can be stripped, reprobed, and subjected to harsh chemical treatments.