Why use pvdf

Before you start your next Western, we can help you decide which membrane will be best for your experiment. In this case, nitrocellulose is more convenient since it does not need to be activated. By activation we mean putting PVDF into methanol, washing it with distilled water and placing it into your transfer buffer. Nitrocellulose was one of the first membranes used and continues to remain a popular choice due to its fast binding properties, affordability, and convenience.

Unlike PVDF, nitrocellulose does not require activation. Ease of hydration makes it an excellent choice for a quick slot blot or Western. Selection of the best membrane depends on the properties of the protein you are trying to detect. Protein size and overall charge are the critical parameters. 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. 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. After all that, having weighed up the pros and cons of each, and after speaking to a number of PhD students from various labs, what appears to play the biggest part in choosing a membrane is personal preference! Preferences which are often influenced or based on what membrane people were first taught with or are available in the lab. So there we have it, nitrocellulose versus PVDF membranes; How have you found different membranes to affect your blots?

Do you have any tips or tricks in general to get the best out of your westerns? PLoS One. Valid application of western blotting. Mol Biol Rep. Epub Feb 8.

PMID : Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. PMID: Western blot: technique, theory, and trouble shooting.