Black ECL blots

[David Young]

In article <dvik-1004951607570001 at bmsb3_micro1.unm.edu>,
dvik at medusa.unm.edu (Dennis Vik) wrote:

> > Has anyone else ever had problems using polyclonal antisera on a western
> > blot detected with ECL resulting in an overall black blot.  It is almost as
> > if the blots were not blocked at all (they were).  The ECL supplier didn't
> > have any suggestions.  If anyone has come across this problem and hopefully
> > a solution, I would appreciate any suggestions. 
> 
Sorry, I didn't see the original post, but I think I have come across this
before. If you are using the Amersham ECL you have to wait a variable
amount of time from the development to the time of exposure. When the
instructions say to do it immediately sometimes you get a black blot, but
if you wait 5-10min then you usually get an okay  blot.

Hope this helps,

David