Northern Blots - can you reuse the buffers and membrane - Even if its dig labelled (Jul/26/2006 )

Hello

I'm running Northern Blots probing for COX2, I cant get these to work but to check that everything else is alright and the RNA is fine then I wanted to reuse the Hybond N+ membrane to check with my B-actin probe. Is this possible or am I wasting my time ?

For the detection method I use DIG labelled probe, that I then bind anti-dig HRP antibody to and use ECL and x-ray film detection.

Could I mix in the two probes at the same time ?

Also when Im blotting and I use lots of chemicals to make up the 20 x SSC, I was wondering if I could just DEPC treat this after and autoclave it and reuse it again and again rather than keep making fresh.

Cheers

Ryan

it's possible.
For stripping membrane, i just add 0.5% boiling SDS and allow to cool.
Membrane is again ready to use. Just wash twice in pre hybridization buffer, and then go for przehybridization step

well if the buffer is used in short period for capillary transfert, why not... (let assume 1week at most).
For >1week use fresh buffer always.

I re-use the 20xSSC for up to five time and no problem.