It is sometimes necessary to detect more than one protein by chemiluminescence on the same Western blot membrane. One way to detect multiple proteins is by stripping and reprobing the membrane with a different primary antibody. Stripping the membrane involves harsh conditions to disrupt the interaction between the membrane-bound protein and the primary antibody. This process enables reprobing with new primary antibody for further protein identification. Although the method of stripping and reprobing risks loss of protein from the membrane, not having to perform a second Western blot does save time and minimizes use of valuable protein samples. Careful consideration of the stripping conditions can help minimize the risk of protein loss from the membrane.
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Which Membrane Should You Use?
Selecting PVDF, Nitrocellulose or Nylon Membranes for Western Blotting | Sino Biological
The invention belongs to the diffusion barrier technical field, be specifically related to the super-hydrophobicization method of modifying on polyvinylidene fluoride film surface. Film distillation at present why also do not have one of large-scale industrial application very chief reason be that to be used for the film permeation flux of film distillation low, the competitiveness of comparing with other isolation technics is not strong. Another is fenestra wetting in running, and flux is stable inadequately. So the requirement that film is distilled with film is to obtain stable high flux. Mohamed Khayet[Khayet M. Pore-size distribution is narrow as much as possible;.
Stripping and Reprobing a Western Blot Membrane
Two membranes types are used in Western blotting applications: nitrocellulose and polyvinylidene fluoride PVDF. How do you choose which one to use? Transfer Options Wet tank vs. PVDF membranes have a higher protein binding capacity than nitrocellulose. Although PVDF has a higher binding capacity, it could result in increased background in some circumstances.
Nitrocellulose or PVDF membrane for western blot Which is better or in what cases you use one or the other???? I read Nitrocellulose is compatible with all standard detection methods and has the best binding capacity without background interference. So, when should you use PVDF? I think, PVDF-membrane is preferred for stripping, I am not sure if you can strip and reprobe nitrocellulose membranes so easily.