Stripping Membranes For Reprobing

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. These considerations include using combinations of detergents, reducing agents, heat, and high or low pH.