[Histonet] IHC protocol for 5 year old Pap smear slides

[hao zhang]

Hi, everyone

I want to do pan-keratin IHC for some five year old pap smear slides. After
I remove the coverslip then I used following protocol for destaining (xylene
5 min, two time change; 3% acetic alcohol (95%) overnight; 70% Etoh for 5
min) and then I put it in PBS, blocking, Prmary Ab, Secondary Ab and DAB.
Problem is that I got less or nothing staining for pan-keratin Ab (even I
used different dilution of Ab). Does anyone know how to recover the antigen
activity for these slides? Any suggest will be apprecaited.--hao