🚀 go-pugleaf

>Hey Folks -- We are trying to immuno-label 20um frozen sections of fixed
>liver through the entire section thickness.  We have tried long (up to
>overnight) incubation with primary antibody, included saponin or triton
>(to 0.2%) and still have problems getting the antibodies into the middle
>of the sections.  Does anyone have any protocols that they have used
>successfully for similar specimens?  The tissue was embedded in
>PVP-Sucrose prior to sectioning, and the sections can be either mounted
>on slides or free-floating.  Please respond directly to me at
>gmartin@welchlink.welch.jhu.edu.   Thanks in advance,
>                                                Greg Martin
>                                                Cell Biology and Anatomy
>                                                Johns Hopkins School of Med.
>

Hi,

We are also limited by the same problem, which I know is not a new one.
However, if anyone is aware of any new tricks that facilitate the
penetration of immuno-label, I would hope that they will post them to the
entire list since this would be of general interest.

Thanks.

Les Malmgren

Leslie T. Malmgren, Ph.D.
SUNY Health Science Center
750 East Adams St.
Syracuse, N.Y. 13210
(315)464-7254