Re: anti-calretinin and pale brown DAB reaction

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From:Debbie Pepperal <>
To:Tibor Ric <>
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 Hi Tibor

We have used the Swant antibody. We microwave antigen retrieved in citrate
buffer pH 6.0 for 10 mins . Dilution of antibody 1:1000 in TBT 
overnight @ 4'C, and ABC as the detection  and DAB as the chromagen. Worked
beautifully. Hope this helps.

Zenobia Haffajee
HAPS, NSW, Australia

At 08:48 PM 12/04/00 +0200, you 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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