Methods to keep stripped blots - (Apr/29/2008 )

How do you guys keep the stripped blots usually? Is there any different keeping it for a long or short term?

I was thinking of just putting the blots in the blocking buffer?

And, is it better to use harsh stripping buffer or mild one? Any difference? I know some antibodies need harsh stripping buffer and doesnt work with mild one. any opinion and why is it so?

thanks!

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If I am storing a stripped blot like over the weekend I put it in TBST or maybe my blocking buffer- both are fine. For longer storage could you dry your membrane? I would worry about things growing in the solution if you stored the membrane wet for very long. As far as stripping buffers go- some antibodies are just harder to release from the membrane so they need harsher conditions. I use the Pierce stripping buffer and it works well- I just adjust time and temperature depending on the antibody.

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thanks for the reply!

Can I just dry it just like that or should I use some vaccuum pump to dry it? And then to use it back again, I have to activate it via methanol right? how does this methanol works actually? i am just curious.

thanks

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you can just air dry the blot.

you have to activate with methanol if your membrane is pvdf. if it is nitrocellulose or nylon then methanol activation is not necessary.

activation of pvdf is necessary because pvdf is hydrophobic and repels water unless it is first "wet" (made a little more hydrophilic) with methanol.

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