Running a Western blot might require several hours of your time, especially when you have multiple target protein molecules.
In this post, we discuss ways of maximizing your Western blot membrane use by stripping and reprobing with another primary antibody. You can use this technique to detect more than one protein molecule on the same membrane, to conserve resources, reduce cost and save valuable time.
Stripping a Western Blotting Membrane
Stripping the Western blotting membrane allows you to use it multiple times for the same set of samples. Whichever membrane stripping method you use, you can remove the primary and secondary antibodies and probe the membrane again.
Multiple uses of your blotting membrane can be especially useful if your proteins of interest are only available in limited quantities. For example, this might be because your protein molecule is difficult to purify, or only present at low concentrations in cell extracts. Or perhaps, you may only have a limited amount of cell extract. Regardless of the reason, you might only have enough protein for one Western blot. On the other hand, your experiment might depend on visualizing two protein molecules of similar molecular weight or a loading control.
For a detailed look at stripping and reprobing methods, see our in-depth article on the subject, or if you are looking for some quick hints and tips and an easy how-to guide then read on below!