Re: anti-calretinin and pale brown DAB reaction

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From:Mary Latimer <>
To:Tibor Ric <>
Content-Type:TEXT/PLAIN; charset=US-ASCII

Dear Tibor you might try to increase the concentration of the igg...I take
it that you are doing an indirect PAP reaction....the alternative is to
repeat the igg and pap steps twice in sequence before reacting it with
DAB hope this helps .....Regards Mary Latimer,Dept of Psychology
University of St- Andrews

On Wed, 12 Apr 2000, Tibor Ric wrote:

> Hi everybody!
> I'm using anti- calretinin from Swant (Code No 7699/4, polyclonal) in my
> project. The problem is that I'm getting too pale DAB reaction. I did
> everything like before for HBME-1 and CK 5/6 and they were not that pale
> brown. I'm sure that the DAB solution is okay. The material I'm using is
> from 1991 to 1999.
> One problem I consider here would be the fixation time which I don't know
> for those older blocks.
> Please, come with suggestions and good ideas for a beginner...
> Today we tried a new antibody from Novocastra Laboratories (clone 5A5,
> monoclonal) and I'm still having that pale brown color...
> The positive controls were pale too.
> Thanks in advance!
> tibor ric
> a Swedish student from the University of Karlstad, Sweden

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