20 March 2015 12 5K Report

Hello All,

Is it possible to re-developing signal without stripping in Western blot? and How do you do it? Please share. It has been stuck in my head for a while. My objective is just to re-develop signal the same protein on the membrane again, in case ECL expired or want to capture image again. No need to change primary and secondary antibody.

From what I read and heard, there are three ways to re-developing signal in western blot:

1. Stripping, blocking, and then re-probing primary and secondary antibodies again, then re-developing with ECL -> lost some protein sample. This method is written in every protocol.

2. Don't strip, just re-probing with secondary antibody, then re-developing with ECL.

3. Washing membrane in TBST for a few time, then re-developing with ECL. -> heard from lab mate -> haven't found reference in google.

Have you ever tried 2nd and 3rd method? Does it work? Please verify

Thank you very much,

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