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Storage of Membrane before addition of primary antibody


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#1 2xzwei

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Posted 08 October 2012 - 01:14 AM

Hi!
I'm not sure how and how long it is possible to store a PVDF membane after protein transfer but before addition of the primary antibody.

The problem is that I've ordered a new antibody and delivery will take some days. So is it possible to do the blot/transfer in advance, store the membrane and then add the primary antibody few days later? If yes, what are the best storage conditions? I've read to keep a membrane in blocking solution at 4 °C but I also found the information that extensive blocking is not recommended...

Does anyone have some tips?

Edited by 2xzwei, 08 October 2012 - 01:15 AM.


#2 gmcstay

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Posted 08 October 2012 - 06:30 AM

Hello,

after the transfer, just keep the PVDF membrane dry, then when the antibody arrives, wet the the membrane with methanol, wash a few times in your western blotting wash buffer (TBS-Tween or similar) then block as you would be doing a normal immunoblot. I do this all the time, and membranes can be stored indefinitely in this manner.

Gavin

#3 2xzwei

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Posted 08 October 2012 - 11:03 PM

Thanks for this suggestion! How to dry the membrane correctly - rinsing with ethanol and then air dry? Should I store it at 4 °C or -20 °C?

#4 cbf88

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Posted 12 October 2012 - 11:39 PM

honestly you can probably just block it then store it in tbs or pbs at 4c for the few days you need, I've done it before just fine (actually just kept it in blocking buffer)

Edited by cbf88, 12 October 2012 - 11:39 PM.





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