PVDF--negative image??

stebby at welchlink.welch.jhu.edu stebby at welchlink.welch.jhu.edu
Thu Aug 20 18:52:00 EST 1998


Help...please!

A friend just called to ask me to troubleshoot a commercially purchased
western blot on PVDF which, when stained for ECL, yielded clear zones where
the proteins in each lane are located on the blot.  No signal was detected,
rather they had what amounted to a negative image of what a coomassie stained
gel would have looked like. It sounds like the protein bands were somehow not
accessible to the antibodies.  I don't use PVDF so I'm stumped already!  Any
takers??

FYI--blots made in their lab onto PVDF (I don't know which supplier) were
used to determine the optimal primary and secondary antibody dilutions prior
to their use of the commercially obtained blot.  The know the antibodies work
and the antigen should be there.

Thanks to all!

Steve Dahl
JHMI

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