Re: Pretreatment for in situ hybridization
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
perfectly.
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Mikael Niku URL: www.helsinki.fi/~mniku/
University of Helsinki Dept. Basic Veterinary Sciences
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- Gandhi
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