How many times can a western blot be stripped?

These days, PVDF membrane is typically used for western blots, allowing the user to strip the blot and then use it repeatedly to probe various proteins.

The basic goal of stripping the blot is to eliminate the primary antibodies from earlier probing. There are several methods for doing this, such as using specific buffers or even heating it in boiling water.

From personal experience, I can tell you that stripping the blot does not always result in the complete removal of the prior antibodies. Additionally, it is important to keep in mind that losing some of the protein that has been transferred or attached to the membrane during the primary antibody removal process makes it best not to strip the membrane more than once.

If somebody could clarify the response, I would be happy to do so.

Salutations!

A Western blot can typically be stripped and reprobed 2-3 times. However, the maximum number of times depends on the durability of the membrane and the efficiency of the stripping process, without significantly losing signal or damaging the proteins bound to the membrane.

A Western blot can typically be stripped and reprobed multiple times, with the exact number of times depending on various factors such as the stripping protocol used, the sensitivity of the detection method, and the quality of the blot. In general, Western blots can be stripped and reprobed up to 2-4 times without significant loss of signal or increase in background noise. However, excessive stripping and reprobing can lead to degradation of the membrane and loss of signal intensity, so it's essential to optimize the stripping conditions and minimize the number of stripping cycles whenever possible. Additionally, some proteins may be more sensitive to stripping than others, so researchers should consider these factors when planning Western blot experiments.