Best chemiluminescence for W.blotting

Keld Sorensen KeldS at uic.edu
Mon Mar 18 09:14:08 EST 1996


[snip a comment about chemiluminescence substrate and 
background]

Remember the substrate can not "create" background.
The substrate simply "reports" where it has found HRP,
therefore, the background is due to the procedure. A 
more sensitive substrate would (all other things equal)
tend to give higher BG. The solution is simple: dilute
the HRP-conjugate (for SuperSignal I would recommend
the HRP-labeled secondary at 1:40,000, but a recent
posting demonstrated that 1:100,000 was better FOR
THAT PARTICULAR APPLICATION!!).
Another solution: take a look at your blocking reagents,
and try some variations.

keld.





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