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Deparaffinization, tissue incubation over night in 85 % ethanol

Deparaffinization Formalin-fixation

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#1 Stefff

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Posted 28 June 2013 - 11:07 AM

Hi all!

I have a question about the deparaffinization procedure of FFPE tissue. Our lab assistant did the deparaffinization as followed:
- Paraffin melting in the oven (55 ˚C)
- Addition of Xylene, removal of Xylene/ paraffin
- Incubation in Xylene for 30 min
- Incubation in 100 % Ethanol for 30 min
- Incubation in 100 % Ethanol for 30 min
- Incubation in 95 % Ethanol for 30 min

Then, he added 85 % Ethanol and forgot the samples so that they were sitting in 85 % Ethanol overnight at room temperature.
So, what do you think, is the tissue still usable or should I start over again? My aim is not to do histological staining but to extract proteins from the tissue.

Thanks for your help!

Steffi

#2 bob1

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Posted 28 June 2013 - 03:09 PM

The proteins will be extensively crosslinked by the fixation and ethanol should preserve this quite well. I don't think there will be any problems.




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