Re: IgG, IgM on FFPE tissue

From:Amos & Theresa <atbrooks@snet.net>

Hi,
    We do not use any pretreatments on these antibodies. We use LSAB+ for
detection, and run them at 1:4000 for IgG, and 1:2000 for IgM. There are
occasional endogenous background issues, but the signal is usually strong
enough to be seen through the "noise". Alternatively one could use Envision to
eliminate this problem, usually the dilution is a little more concentrated
though.
Amos Brooks

antje.marcantonio@pharma.Novartis.com wrote:

> Hello Histonetters,
>
> we try to do direct immunofluorescence on paraffin sections of monkeys with
> Dako antibodies for IgG and IgM. Until now we had no success with HIER or
> enzyme digestion with trypsin. We'll go for their recommended pronase
> treatment before the staining and we are hoping ....
> But now I would like to know from you experts out there: any
> recommendations? How about procedures for human FFPE tissue ??
> And again thanks a lot in advance for all your comments and ideas.
> Have a great time,
>
> Antje
>
> Antje Marcantonio
> BU Transplantation
> Novartis Pharma AG, Basel




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