It is necessary for me to save my PVDF membranes from western blots in order to reprobe them with different antibodies in the future. Currently, I keep my membranes in 50 mL tubes filled with 1x PBS at 4C. I have also seen membranes stored dry between sheets of plastic at 4C. Do either of these methods affect the fidelity of protein binding or antibody attachment? I feel like the storage in PBS may gradually wash off antibodies or proteins. And I feel like dry storage may affect protein confirmation. How do you store your western blot membranes?
Hi David,
I will prefer you storing you membrane in 4˚C,most especially PVDF has the ability to be stable over a long time of storage than nitrocellulose. it is possible to store a PVDF membrane and preserve sample proteins for extended periods of time by following the steps below.
1.Sandwich the dry PVDF blot between two clean sheets of Whatman 3MM paper.
2.Place the sandwich between two sheets of card stock or thin cardboard.
3.Use paperclips to clip the stack together on the edges.
4. Place the stack in a plastic bag and seal the plastic bag closed.
5. Store the blot at 4 ˚C for up to 2 weeks, -20 ˚C for up to 2 months, or -70 ˚C for longer storage.I hope these suggestions will help you.
All the best!
Hi David,
I use to store my PVDF membrane dry between sheets of plastic at 4C for at least 1 week at 4°C and it work very well and does not affect protein attachment.
Didn't know that they could be stored longer
For 20 plus years, I have dried my PVDF membranes and stored them in lab notebooks in a pocket of kimwipes taped to the page in the notebook where they were run. WE have re-probed them years later with no ill effects. I have seen mold and other problems, even at 4C, storing them wet. especially for extended periods.
Store dry as suggested above due to mold etc. If you wish to probe the membranes again with another set of antibodies....strip before you let the membrane dry out or the antibodies from first probe will remain stuck to the membrane. If your second probing is with antibody raised in different species than first probing antibody, then just incubate for 30 min in pH 2 water made with HCl, then rinse well in water and dry store.....This removes secondary antibody conjugate used in first probing, leaving the way clear to probe with other species primary antibody and secondary against another species
The best way to store PVDF membrane is dry the membrane and then keep at -20oC without folding.
you can cast it in glass subbstrate and store it adhehere or in water bath
you can cast it o glass substrate and store it adhere or in water bath
Dear David,
In order to store PVDF membranes for long time, they should be stacked in non-biodegradable material with a temperature ~ 4 °C. It is worth mentioning that membranes must be totally dried before storing.
- Badges
- Science method