John E. Smith III jes2u at virginia.edu
Sun Feb 4 16:33:58 EST 1996

Kathleen Clark wrote:
>...The ECL kit produces a large number of nonspecific bands that I >can't eliminate using conventional measures (long blocking, dilute >antisera and lowering amount of sample on gel)....

Kathleen, this may be caused by nonspecific binding of the secondary
antibody with proteins in your ovary extract.  You can try 
pre-adsorbing your secondary antibody by incubating with a second blot 
which has not been exposed to primary antibody.  Any antibodies in 
your polyclonal secondary which cross react will be removed, then you
can use this as your secondary for your blot of interest.

Hope this helps.

-John Smith
jes2u at virginia.edu

More information about the Cellbiol mailing list