Chemiluminescence, western blotting, problems...

phillip buckhaults buckhaults at bscr.uga.edu
Tue Mar 5 01:34:15 EST 1996


Giampaolo Minetti wrote:
> 
> Hello,
> I am experiencing problems with Enhanced Chemiluminescence Detection
> (Amersham ECL) and western blotting.
> What happens is that after adding the ragent, incubating for 1 min and
> taking the nitrocellulose into the cassette for exposure (I use to
> cover the membrane with Saran Wrap), everything is OK.
> After 2 or 3 minutes, however, the entire membrane is luminescent. The
> bands I'm interested in are there, but you cannot take a good
> exposure because of the extremely high background.

(much ascii deleted)

Giampaolo,

do you drain your blot well after soaking in the ECL reagent?  try letting all the 
liquid drain that will, touching the edge of the blot to the container to get the 
last bit off.  

anyway, this helped us.

good luck.
-phil



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