What should I do to get optimal transfer results?

Larger proteins necessitate lengthier transfer times and lower voltages, whereas smaller proteins can be transferred faster using higher voltages. Additionally, selecting a buffer that suits the gel, the target protein, and membrane type is essential for successful western blotting. One should always confirm the effectiveness of protein transfer by staining the membrane with a total protein stain or utilizing a pre-stained protein ladder. Additionally, regular maintenance and inspection of the blotting apparatus can help prevent problems such as uneven heating and inconsistent current flow, which may impair transfer efficiency. The choice of membrane can also influence the sensitivity of detection and the ability to bind proteins. Nitrocellulose and polyvinylidene difluoride (PVDF) are two common types of membranes used in Western blotting. Nitrocellulose membranes are popular due to their high capacity to bind proteins and their ability to interact with various detection methods.