To strip or not to strip - that is the question (Sep/25/2006 )

Hi all,

I was wondering, when is it important to strip a membrane before reprobing with a new antibody? Is it only for proteins that are similar in size or are there other reasons? Also, doesn't it take more of a toll on the membrane? I like to get the most probes out of one membrane but until now, I've always stripped the membrane before reprobing. Is it really necessary? Sorry, i know it's kind of an elementary question but I'm still a newbie

If I know that the antibodies are so specific that I will only get one band, and if the different proteins I want to label do not have the same molecular weigh, I do not strip, and sometime I also incubate the two antibodies together (if the second protein is not the double size of the other, because I would not know if I see the big protein or a dimer of the small protein).

i tend to do as misselle, excepting i don't mix primary antibodies

some contributions in the following discussion give some good hints to your question:

http://www.protocol-online.org/forums/inde...showtopic=20518