Re-ECLing nitrocellose??

Paul pd1 at mole.bio.cam.ac.uk
Wed Nov 1 09:18:04 EST 2000



Neal Robert Melvin wrote:

> Has anyone ever tried to 're-do' an ECL reaction on an NC membrane that
> has been previously processed, and subsequently dried?? Do you have to
> strip the membrane first, and re-apply primary and secondary antibodies
> even if you want to re-probe with the same primary as you used
> originally??
>

We do this fairly often: we don't bother stripping but we've found that we

do have to re-apply the primary and secondary Abs.  Dumping fresh ECL
substrate on without fresh Abs doesn't work in our hands, even on a fresh
blot that's not dried out.  Not sure why, but the guess is that the HRP
becomes inactivated.


Cheers

Paul Digard






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