Have you ever wished you could transfer the same SDS-PAGE gel twice? Sometimes, when you are blotting for many different proteins of similar size, stripping and reprobing multiple times can become impractical. Here’s a simple diffusion transfer method that can be used to generate duplicate membranes from a single gel:
- Take a glass plate, or something that is similarly sturdy and flat.
- Layer the following onto the glass plate in order, smoothing each layer out to remove air bubbles:
- Three pieces of blotting paper (e.g. Whatman 3MM) that have been soaked in phosphate-buffered saline (PBS).
- Lay out a pre-soaked membrane (I’ve only tested nitrocellulose, but PVDF should be fine).
- Your gel.
- Another pre-soaked membrane.
- Three more pieces of PBS-soaked blotting paper.
- Add another glass plate.
- Tightly wrap the whole sandwich in plastic wrap.
- Place a heavy object (e.g. a 1L bottle of buffer) on top.
- Go home for the night, or for the weekend.
Try this method out, and let us know how it goes! It sounds too low-tech to work, but the few times that I’ve tried it has given me great results.