Can I reuse a blot that worked previously for a specific antibody and stored at -80 degrees to test for a different antibody? The blot that I am using currently is a nitrocellulose blot.Answers or suggestions would be helpful.
Defintely not a good idea to store nitrocellulose at -80C. I have never done this but I imagine it renders a brittle membrane even more brittle and susceptible to damage
To store s membrane already probed for subsequent reasons probing it is perfectly acceptable - indeed common practice - to wrap in cling film and either store protected at room temp in a cold room or the fridge
before re probing strip the membrane by subjecting to a stripping buffer (plus beta mercaptethanol) at 37 to 60C for up to 1 hour at this elevated temp or alternatively leave in this stripping buffer overnight at 4C
For low level proteins it is even possible to strip the membrane by leaving in the cold room in TBS buffer plus 1% tween 20: thus if I am going to re probe a membrane over the course of 1 week I will simply with such low level proteins leave in TBS + 1 % tween in a cold room for 48 hours (instead of dry in cling film either at room temp or in the fridge); and then subject to stripping at elevated temp as described for about 30 min
Finally having stripped re block for 1 hour then apply your second primary antibody; and then complete the western protocol as normal