Re: Pretreatment.


what is your tissue?  if it is bone or cartilage HIER can be a real problem in
my experience.

Thu-Thao Pham wrote:

> Hi everyone,
> As some of you may know that I post a questions regarding CD31 and
> pretreated the slides with DAKO Target Retrieval Solution. However, my
> tissue got eaten up and the mophology look back. Some tissues even about to
> come off the slides. I try to be very careful when I take the slides out and
> washing the slides.
> I did as followed:
> 1) Deparaffinized (Histoclear to water)
> 2) Washed slides in TBST 3x for 5 min.
> 3) Pretreatment with TRS by steam for 20 min.)
>     - first heat the TRS to boil and let the TRS
>       solution become clear before place slides into
>       the TRS heated solution (95C).
> 4) Remove the jar from steamer and let it cool to room temperature for
> 30min.
> 5) Gently remove the slides and washed in ddH2O 3x for 5 min.
> 6) Washed slides in TBST 3x for 5 min.
> 7) Proceed with 3% H2O2 for 5min.
> 8) Washed TBST
> 9) CD31 diluted in 1% BSA (in TBST).
> 10) Followed by DAKO kit
> After preteated my slides can I leave at room temperature for at least an
> hour to cool down?
> Many thanks if anyone can help.
> Thao
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