Hello, I am trying to optimize my northern hybridization. I realized that the membrane (SensiBlot
Plus Nylon Membrane) that we are using has reached it's expiration date many years ago. Maybe somebody could explain how exactly it can affect my experiment/samples and why? Thank you very much.
Hard to believe how properly stored membranes can expire. It may be just a sales pitch. We have no trouble in using old membranes for western blotting. If you are concerned then try parallel expts with expired and unexpired membranes and see if their is any difference in signal. Regardless of results you can write a small technical note for the benefit of others!!!
You can use methylene blue to stain your northern membrane post transfer to assess transfer efficiency, but I have no experience using membranes after their expiration. Good luck!
Hard to believe how properly stored membranes can expire. It may be just a sales pitch. We have no trouble in using old membranes for western blotting. If you are concerned then try parallel expts with expired and unexpired membranes and see if their is any difference in signal. Regardless of results you can write a small technical note for the benefit of others!!!
- If packs of membranes are many years old they tend to go brittle and sometimes even discolour slightly (due to photo oxidation):
- I have seen this with Old Nitrocellulose
- If it is excessively old and brittle you might find proteins don't transfer as well (they tend not to stick) and also blocking doesn't work as well so you get more general background
- I would say that is your membranes are only 1 year out of date and not excessively brittle they will be fine
- If however they are say 5 years out of date, slightly yellow and brittle do not use (for reasons given)
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