colony screening with Amersham ECL (RPN 3000 and 3004)

bhjelle at carina.unm.edu bhjelle at carina.unm.edu
Fri Dec 18 09:53:55 EST 1992


In article <1992Dec16.151837 at genethon.fr> bouchier at genethon.fr (Christiane Bouchier) writes:
>
>A week ago I asked whether anyone had ever carried out tests for screening
>colonies  with Amersham ECL direct system. So far I received no suggestions
>that could help me overcome the problem we have of detecting peroxydase 
>specific hybridization because of a high background.
>Once again thanks for your help !
>
For months we used Amersham ECL for a dot blot assay that is
used a lot in my lab. Backgrounds were highly variable but
eventually got so out of hand we had to switch back to
32P. We were unable to fix the problem with the blocking
procedures recommended by Amersham or any other obvious
manipulation. The problem was nonspecific blobs of signal
that covered parts of the membrane that had never seen
any biological material.

Amersham will have suggestions that you might want to
try. Contact their rep. However, we found it easier to
switch back to isotopes than to do Amersham's R&D.

Brian



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