Currently I'm using "Restore plus western blot Stripping Buffer" for reprobing a new antibody on my blots. I am not that happy with the results because the residues of first use are still there!
I do the classic stripping protocol with beta-mercaptoethanol and SDS and it works fine as long as you do it for a short period of time.
First, what kind of antibody are you trying to strip off? Certain ones, especially for phosphorylated proteins don't like to come off. You might as well run a second gel for total protein. Second, I am not familiar with the stripping solution you are using. How long do you leave it on your membrane? What temperature? All these details are important. The higher the antibody affinity, the harder it is to strip off. Pierce's stripping solution is pretty good. Be careful when using these kind of solutions, as they impact antigens in your membrane, which impacts the secondary detection you are hoping to perform.
Restore plus western blot Stripping Buffer, works pretty good for me! here are some suggestions for using this buffer:
1- wash your membrane with PBS or TBS several times before you add the Restore buffer.
2- At room temperature, you should strip your membrane for at least 30 min; at 37 c, 15 mi.
3- sometimes you need to strip twice (each 15 min); refresh the Restore buffer after 15 min.
Good luck
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