Storing Western blot membranes - What should I do after detection and stripping? (Dec/11/2007 )
I have a membrane which I need to strip and re-probe but we have run out of antibody. Obviously I don't want to have to run the whole experiment again after xmas so I need to store the membrane somehow.
I have heard that you can dry the blots out and then simply re-wet using methanol, but has anyone got any actual evidence for this?
I'm using PVDF membranes if it is any help!
I've been storing PVDF membrane for few days after blocking with TBS-Tween, wrapped in saran. No re-wetting with methanol was necessary, just washed it for few minutes with TBS-Tween.
It was not my idea to be honest, I got this advice from here (sorry, I've bad memory for names)
we keep the membrane between the 2 layer of transparent copy paper (as you would go for developing) and wrap this in plastic strech film and store in the fridge. For reusing just wash in PBST/TBST (what ever you are using).
Hi, Rosie. We use to keep the membranes frozen until we need them. After transfer, I just put them in methanol for a few seconds to push all the water away, dry them with a hair-drier (just cold air stream) and wrap them individually in kitchen foil. I store them at -20ºC, and they work ok, some of the membranes I've recently used have been stored for more than 6 months. When you need one of them, just take it away from the envelope, activate it with methanol, block as you usually do, and it's ready to continue with your assay. Ah! You can also keep then blocked, I don't do it but I know someone here does and it works. The trick is to completely dry the membrane before freeze (you can get more info about drying the membranes in Millipore page).
I also routinely dry out my PVDF membranes to be blotted again at a later date. I store mine tightly sealed at 4°C which seems to work for a week or so, but maybe for longer-term storage you might wish to go colder.