Re: Pretreatment for in situ hybridization

From:Phyllis Davie

Hi Christina,
    We are using heat for the EBER1 probe.  We pretreat in 10mM citrate, pH
6.0 in a microwave pressure cooker for 15 minutes at pressure.  We did
comparisons between the above method and Proteinase K and Pepsin.  The HIER
was far superior.  I don't know about B-5, or the other probes, we haven't
tried them.

Best of luck,

Phyllis Davie
PhenoPath Laboratories--Seattle, WA

on 6/6/02 8:35 PM, C Sawrenko at 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?  We are attempting ISH for kappa, lambda and EBER.
> Also has anyone had any success on B5 fixed tissue for the above?
> Thanks in advance for your help!
> Christina Sawrenko
> Histopathology
> BC Cancer Agency
> Vancouver, BC

<< Previous Message | Next Message >>