ChemiGlow Re-ECLing nitrocellose??

WSchick at WSchick at
Thu Nov 2 18:52:21 EST 2000

AlphaInnotech demoed their FluorChem ChemiImager, and simply added their 
ChemiGlow HRP substrate to an ECL blot without rinsing or reapplying 
antibodies.  Worked fine with more light output for faster imaging and lasted 
longer so their Movie mode could take different exposures automatically.   
Some bands were so bright, we plan to test their recommendation to dilute 
their substrate.

They list this on their website,

Walt Schick

> Thats strange, In my lab we currently are experiencing problems with the
>  ECL.
>  It gives rise to "clouds" on the film so what people do when they see
>  this is to  simply rinse the blotted PVDF-membrane in dd-water or PBS
>  and re-apply ECL. After that the blot is fine. No clouds. But mind you,
>  this is on wet - non-frozen blots.
>  -Max
>  Paul wrote:
>  > 
>  > 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 
>  > 
>  > 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
>  > 


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