Recently we have been having issues re-probing PVDF membranes and have made and bought fresh antibodies, yet still see poor labeling even on the first re-probing. We use ethanol over methanol for safety and previous lab members supposedly found no difference when using ethanol.
Would anyone know if methanol is better than ethanol for PVDF activation and if it will improve transfer if used in transfer buffer, compared to ethanol?
Hello Kevin,
You can use ethanol to activate the PVDF membrane instead of methanol.
But there are two reasons why methanol is commonly used to activate the PVDF membrane.
1. If you use ethanol to activate the PVDF membrane, it must be of high quality. If impurities and solvents are present in ethanol, it can mess up with your transfer process. Procuring pure ethanol would not be an easy task. You will have to spend more to obtain ethanol of high-quality grade.
2. Using ethanol may require extra minutes to activate the membrane. If you use the water step, you will need more time to equilibrate the membrane with water. Ethanol/water mixtures are more viscous than methanol and it will take longer for ethanol to diffuse out of the membrane.
Moreover, ethanol is more hydrophobic than methanol because methanol is a small solute having a single hydrophilic hydroxyl group whereas ethanol possesses an additional hydrophobic carboxyl group. So, you will also need to equilibrate with the transfer buffer for a longer time.
I would prefer methanol because it is more effective at activating the membrane and facilitating protein binding.
Best,
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