How many times can a western blot be stripped?
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Personally, I strip three times very gently and my signals are very uniform. I notice that if I probe my blots out of sequence (going from, say, actin, strip, tubulin to tubulin, strip, actin) I can get high variation.
As long as it's on a shaker the stripping across lanes should be equal, since you should be putting on an ample amount of stripping buffer.
Hard to say how much protein is left since that depends so heavily on the efficiency of your transfer and stripping conditions. I've had some antibodies that are so well-made that trying to remove them nearly takes the ladder with.
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