Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
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:
Areas of PVDF coated with transferred protein are capable of wetting out in 20% methanol while the surrounding areas of PVDF are not.
Possible Cause
Remedy
Inappropriate membrane
Transillumination works best with Immobilon®-P transfer membrane. It is not recommended for nitrocellulose or Immobilon®-PSQ transfer membrane.
Membrane wasn’t completely dried prior to wetting with methanol
Be sure that the membrane was dried completely after the transfer prior to immersing it in the 20% methanol solution. Make sure to use a 20% methanol solution.
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.
Air bubbles
The blot should not have any air bubbles on the surface. Gently pull the membrane across the edge of the container to remove bubbles.
