I have a problem where I used different protocol (Ponceau, acid wash (with SDS and low pH) and one commercial) for stripping followed by re-probing with new antibodies that had the same secondary antibody as the initial blotting. I always get residual signal coming from the first antibody.
Can anyone recommend a robust assay for stripping (homemade or commercial) that has been tested to work for stripping of the primary antibody (secondary is insufficient in my case)?
Hi Peter! From my experience there will be always faint residuals of the previous antibody if it is too strong (i.e. GAPDH). however, I always get good stripping results with ReBlot Plus Strong Antibody Stripping Solution, 10x (millipore). You may want to try it, but dont over do it (max 30 min with shaking) or your membranes will lose their proteins as well.
Good luck,
Laila :)
Thanks laila I saw this reblot mix add and didn't know if it worked well.. I will try it out now.
Cheers
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