Pretreatment.

From:Thu-Thao Pham

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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