When it comes to western blotting, choosing the best membrane for your application can mean the difference between crystal clear bands and a blotchy mess. There are four key criteria you should consider when choosing a western blotting membrane.
1. Protein binding capacity – PVDF has a higher protein bind capacity than nitrocellulose. This means that PVDF is more sensitive but also more proton to background noise. Choose PVDF for applications requiring high sensitivity.
2. Pore size – nitrocellulose and PVDF are available in a variety of pore size ranging from 0.45 µm or 01̶ 02 µm. Most protein applications are suitable for 0.45 µm pore sizes; however, 0.1 µm is recommended for protein or peptides less than 15 kDa in size.
3. Durability – nitrocellulose is more fragile than PVDF. Making PVDF ideal for experiments requiring stripping and reprobing.
4. Autofluorescence – Some membranes are more autofluorescence more than others. PVDF is more susceptible fluorescence than other membrane types, so nitrocellulose or low fluorescent PVDF are more ideal for fluorescence western blotting experiments.
There are numerous factors when selecting the right membrane for your western blotting experiments. Knowing how each parameter effects each result will help you know which one to use.
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