How many times can one strip a membrane? - (Jan/08/2007 )

Hi All.
What is the max no. of times you have stripped a nitrocellulose membrane?
What was the amount protein loaded?

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I was loading 20 to 30 µg and stripping twice (3 blottings)
But it depends on the amount of your protein of interest and on the affinity of your antibody.
If you have an good antibody, and a protein well expressed, I assume you can strip more. the most important is to always blot first the weaker signal.

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I have experience with PVDF; there are differences for 1D and 2D blots; I have once stripped a 2D blot the 7th time and it is still to use (no background from former Ab´s but specific immunosignals); for 1D blots I do not strip more than 3 times

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