Related Resources: Brochures | Application NotesMost Western blot users visualize transferred proteins using stains, such as Ponceau S or Coomassie® Blue. However, users of PVDF membrane have the opportunity to assess transfer efficiency using transillumination. Areas of PVDF coated with transferred protein are capable of wetting out in 20% methanol while the surrounding areas of PVDF are not. In the areas where the PVDF wets, it becomes optically transparent, allowing visualization of protein bands using backlighting and photographic archiving.
Click on the protein visualization topics to read about the possible causes and remedies:
Use sufficient volume of incubation solutions and ensure that the membrane is completely covered with these solutions during incubation.
The container used should be large enough to allow solution to move freely across the blot. Do not incubate more than one blot at a time in that same container. In addition, the protein side of the blot should be facing up so as not to be interacting with the bottom surface of the container.
The blot should not have any air bubbles on the surface. Gently pull the membrane across the edge of the container to remove bubbles.