I've been doing immunos close to 20 years and did not encounter this problem
until HIER came into play. It has something to do with heat retrieval with
particular antibodies ( I have a problem with
Vimentin and cytokeratin, clone 34 BE12). If anyone has any suggestions for
reason and possible remedy, I'm sure we'd all be grateful.
----- Original Message -----
From: "Coskran, Timothy M"
Sent: Thursday, December 12, 2002 2:29 PM
> Has anyone ever encountered the following type of IHC background problem:
> With two separate antibodies, one a cytoplasmic marker and the other a
> membrane marker, every single nucleus in the section turns brown. The
> itself works, as most of the time you can read through the background, but
> all test slides and negative controls (omission of primary) are affected.
> Both antibodies are rabbit polyclonals, both are steam retrieved one with
> citrate pH 6 the other with EDTA. Both are detected using a whole IgG
> biotinylated goat anti rabbit secondary followed by Elite ABC and Dako
> Liquid DAB+. Blocks included 3.0% hydrogen peroxide (aq), Dako Biotin
> Blocking system and Dako Protein block. All were stained on an IHC
> In addition, both of theses antibodies have been reliable for months
> the above protocol steps), until now......
> Tim Coskran
> LEGAL NOTICE
> Unless expressly stated otherwise, this message is confidential and may be
privileged. It is intended for the addressee(s) only. Access to this E-mail
by anyone else is unauthorized. If you are not an addressee, any disclosure
or copying of the contents of this E-mail or any action taken (or not taken)
in reliance on it is unauthorized and may be unlawful. If you are not an
addressee, please inform the sender immediately.
<< Previous Message | Next Message >>