Re: Pretreatment for in situ hybridization

From:Mikael Niku

C Sawrenko wrote:
> Hello Histonetters,
> Does anyone out there have any experience using heat (by steam, microwave or
> pressure cooker) instead of enzyme (Proteinase K) for pretreatment prior to
> in situ hybridization?  

I'm routinely using microwave heating (2xSSC buffer pH6,
500 ml, 750W, 15 min, no pressure) for paraformaldehyde-
fixed sections. For genomic ISH I'm doing, this usually
needs to be combined with protease treatment. Without
the heating PFA-fixed samples are almost useless for
this staining, with heating + protease they work

    Mikael Niku             URL:
    University of Helsinki  Dept. Basic Veterinary Sciences
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